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The Citron homology domain of MAP4Ks improves outcomes of traumatic brain injury
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作者 Xiaoling Zhong Wenjiao Tai +4 位作者 Meng-Lu Liu Shuaipeng Ma Tianjin Shen Yuhua Zou Chun-Li Zhang 《Neural Regeneration Research》 SCIE CAS 2025年第11期3233-3244,共12页
The mitogen-activated protein kinase kinase kinase kinases(MAP4Ks)signaling pathway plays a pivotal role in axonal regrowth and neuronal degeneration following insults.Whether targeting this pathway is beneficial to b... The mitogen-activated protein kinase kinase kinase kinases(MAP4Ks)signaling pathway plays a pivotal role in axonal regrowth and neuronal degeneration following insults.Whether targeting this pathway is beneficial to brain injury remains unclear.In this study,we showed that adeno-associated virus-delivery of the Citron homology domain of MAP4Ks effectively reduces traumatic brain injury-induced reactive gliosis,tauopathy,lesion size,and behavioral deficits.Pharmacological inhibition of MAP4Ks replicated the ameliorative effects observed with expression of the Citron homology domain.Mechanistically,the Citron homology domain acted as a dominant-negative mutant,impeding MAP4K-mediated phosphorylation of the dishevelled proteins and thereby controlling the Wnt/β-catenin pathway.These findings implicate a therapeutic potential of targeting MAP4Ks to alleviate the detrimental effects of traumatic brain injury. 展开更多
关键词 adeno-associated virus Citron homology Citron homology domain gene therapy mitogen-activated protein kinase kinase kinase kinases traumatic brain injury
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Designing and Cloning of the Gene Sequence Encoding Silk Fibroin Amorphous Domain 被引量:3
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作者 黄海燕 田智芳 +2 位作者 裔洪根 杨云星 王建南 《Journal of Donghua University(English Edition)》 EI CAS 2012年第6期489-492,共4页
To provide materials used in investigating the relationship between amino acid compositions of silk-like protein, structure, and functions, especially the biological functions, the motif genes encoding the silk fibroi... To provide materials used in investigating the relationship between amino acid compositions of silk-like protein, structure, and functions, especially the biological functions, the motif genes encoding the silk fibroin amorphous domain, SGFGPVANGGSGEASSESDFGSSGFGPVANASSGEASSESDFAG(F) were designed and extended using a "head-to-tail" construction strategy. The designed genes were cloned into PSLFA1180FA and multimerized to form structures containing a two-timer, a four-timer, an eight-timer, and a twelve-timer. All the resulting plasmids were digested using the restriction enzyme BamHI and the double-enzymes BglII/HindIII. Restriction enzyme analysis and DNA sequencing revealed the motif was successfully cloned into PSLFA1180FA and multimerized to form a twelve-timer without gene deletion or mutation. 展开更多
关键词 Escherichia coli silk fibroin amorphous domain gene cloning DNA electrophoresis
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Expression of Major Antigen Domains of E2 Gene of CSFV and Analysis of its Immunological Activity 被引量:1
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作者 Hong TIAN Xiang-tao LIU Jing-yan WU You-jun SHANG Tao JIANG Hai-xue ZHENG Qing-ge XIE 《Virologica Sinica》 SCIE CAS CSCD 2008年第4期247-254,共8页
E2 is an envelope glycoprotein of Classical swine fever virus (CSFV) and contains sequential neutralizing epitopes to induce virus-neutralizing antibodies and mount protective immunity in the natural host. In this stu... E2 is an envelope glycoprotein of Classical swine fever virus (CSFV) and contains sequential neutralizing epitopes to induce virus-neutralizing antibodies and mount protective immunity in the natural host. In this study, four antigen domains (ABCD) of the E2 gene was cloned from CSFV Shimen strain into the retroviral vector pBABE puro and expressed in eukaryotic cell (PK15) by an retroviral gene expression system, and the activity of recombinant E2 protein to induce immune responses was evaluated in rabbits. The results indicated that recombinant E2 protein can be recognized by fluorescence antibodies of CSFV and CSFV positive serum (Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China) using Western blot, indirect immunofluorescence antibody test (IFAT) and ELISA, Furthermore, anti-CSFV specific antibodies and lymphocyte proliferation were elicited and increased by recombinant protein after vaccination. In the challenge test, all of rabbits vaccinated with recombinant protein and Chinese vaccine strain (C-strain) were fully protected from a rabbit spleen virus challenge. These results indicated that a retroviral-based epitope-vaccine carrying the major antigen domains of E2 is able to induce high level of epitope-specific antibodies and exhibits similar protective capability with that induced by the C-strain, and encourages further work towards the development of a vaccine against CSFV infection. 展开更多
关键词 Classical swine fever virus (CSFV) E2 gene Antigen domains Retroviral vector Immunologicalactivity
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Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants
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作者 Pavel Cejnar Ludmila Ohnoutkova +2 位作者 Jan Ripl Tomas Vlcko Jiban Kumar Kundu 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第11期2492-2500,共9页
Wheat dwarf virus (WDV), an important cereal pathogen, is closely related to Maize streak virus (MSV), a model virus of the Mastrevirus genus. Based on its similarity to known MSV resistance strategies, a truncate... Wheat dwarf virus (WDV), an important cereal pathogen, is closely related to Maize streak virus (MSV), a model virus of the Mastrevirus genus. Based on its similarity to known MSV resistance strategies, a truncated part of the WDV replication- associated (RepA) gene (WDVRepA215) and the WDV RepA gene with a mutated retinoblastoma-related protein (RBR) interaction domain (WDVRepA215RBRre^t) were cloned into the plPKb002 expression vector and transformed into immature embryos of spring barley cv. Golden Promise plants through Agrobacterium-mediated transformation. A detailed study of T1-generation plants infected by leafhoppers (Psammotettix alienus) fed on infection sources of variable strength was performed over a 5-week period encompassing the initial stages of virus infection. A DNA WDV TaqMan qPCR assay normalized using the DNA puroindoline-b SYBR Green qPCR assay for samples on a per week basis revealed an approximately 2-week delay in WDVRepA215RBR^mut plants to WDVRepA215 plants before significant increases in the WDV viral levels occurred. Both WDVRepA215 and WDVRepA215RBR^mut plants showed similar levels of transgenic transcripts over the screened period; however, the transgenic plants also showed increased numbers of infected plants compared to the control plants. 展开更多
关键词 Wheat dwarf virus (WDV) truncated Rep gene RBR domain qPCR screening
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Two memory associated genes regulated by amyloid precursor protein intracellular domain Novel insights into the pathogenesis of learning and memory impairment in Alzheimer's disease
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作者 Chuandong Zheng Xi Gu Zhimei Zhong Rui Zhu Tianming Gao Fang Wang 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第5期341-346,共6页
In this study, we employed chromatin immunoprecipitation, a useful method for studying the locations of transcription factors bound to specific DNA regions in specific cells, to investigate amyloid precursor protein i... In this study, we employed chromatin immunoprecipitation, a useful method for studying the locations of transcription factors bound to specific DNA regions in specific cells, to investigate amyloid precursor protein intracellular domain binding sites in chromatin DNA from hippocampal neurons of rats, and to screen out five putative genes associated with the learning and memory functions. The promoter regions of the calcium/calmodulin-dependent protein kinase II alpha and glutamate receptor-2 genes were amplified by PCR from DNA products immunoprecipitated by amyloid precursor protein intracellular domain. An electrophoretic mobility shift assay and western blot analysis suggested that the promoter regions of these two genes associated with learning and memory were bound by amyloid precursor protein intracellular domain (in complex form). Our experimental findings indicate that the amyloid precursor protein intracellular domain is involved in the transcriptional regulation of learning- and memory-associated genes in hippocampal neurons. These data may provide new insights into the molecular mechanism underlying the symptoms of progressive memory loss in Alzheimer's disease. 展开更多
关键词 Alzheimer's disease amyloid precursor protein amyloid precursor protein intracellular domain chromatin immunoprecipitation gene regulation chromatin DNA
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Assessment of pathogenicity and functional characterization of APPL1 gene mutations in diabetic patients
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作者 Ping Shi Yang Tian +7 位作者 Feng Xu Lu-Na Liu Wan-Hong Wu Ying-Zhou Shi An-Qi Dai Hang-Yu Fang Kun-Xia Li Chao Xu 《World Journal of Diabetes》 SCIE 2024年第2期275-286,共12页
BACKGROUND Adaptor protein,phosphotyrosine interacting with PH domain and leucine zipper 1(APPL1)plays a crucial role in regulating insulin signaling and glucose metabolism.Mutations in the APPL1 gene have been associ... BACKGROUND Adaptor protein,phosphotyrosine interacting with PH domain and leucine zipper 1(APPL1)plays a crucial role in regulating insulin signaling and glucose metabolism.Mutations in the APPL1 gene have been associated with the development of maturity-onset diabetes of the young type 14(MODY14).Currently,only two mutations[c.1655T>A(p.Leu552*)and c.281G>A p.(Asp94Asn)]have been identified in association with this disease.Given the limited understanding of MODY14,it is imperative to identify additional cases and carry out comprehensive research on MODY14 and APPL1 mutations.AIM To assess the pathogenicity of APPL1 gene mutations in diabetic patients and to characterize the functional role of the APPL1 domain.METHODS Patients exhibiting clinical signs and a medical history suggestive of MODY were screened for the study.Whole exome sequencing was performed on the patients as well as their family members.The pathogenicity of the identified APPL1 variants was predicted on the basis of bioinformatics analysis.In addition,the pathogenicity of the novel APPL1 variant was preliminarily evaluated through in vitro functional experiments.Finally,the impact of these variants on APPL1 protein expression and the insulin pathway were assessed,and the potential mechanism underlying the interaction between the APPL1 protein and the insulin receptor was further explored.RESULTS A total of five novel mutations were identified,including four missense mutations(Asp632Tyr,Arg633His,Arg532Gln,and Ile642Met)and one intronic mutation(1153-16A>T).Pathogenicity prediction analysis revealed that the Arg532Gln was pathogenic across all predictions.The Asp632Tyr and Arg633His variants also had pathogenicity based on MutationTaster.In addition,multiple alignment of amino acid sequences showed that the Arg532Gln,Asp632Tyr,and Arg633His variants were conserved across different species.Moreover,in in vitro functional experiments,both the c.1894G>T(at Asp632Tyr)and c.1595G>A(at Arg532Gln)mutations were found to downregulate the expression of APPL1 on both protein and mRNA levels,indicating their pathogenic nature.Therefore,based on the patient’s clinical and family history,combined with the results from bioinformatics analysis and functional experiment,the c.1894G>T(at Asp632Tyr)and c.1595G>A(at Arg532Gln)mutations were classified as pathogenic mutations.Importantly,all these mutations were located within the phosphotyrosinebinding domain of APPL1,which plays a critical role in the insulin sensitization effect.CONCLUSION This study provided new insights into the pathogenicity of APPL1 gene mutations in diabetes and revealed a potential target for the diagnosis and treatment of the disease. 展开更多
关键词 Adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 1 Maturity-onset diabetes of the young Bioinformatics analysis gene mutation domain
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Modeling the Structure of Yeast MAT<i>α</i>1: An HMG-Box Motif with a C-Terminal Helical Extension
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作者 Doba Jackson Tarnisha Lawson +1 位作者 Robert Villafane Lisa Gary 《Open Journal of Biophysics》 2013年第1期1-12,共12页
The yeast MATα1 is required for the activation of α-specific genes in Saccharomyces cerevisiae and thus confers the α-cell identity of the yeast. MATα1 contains a domain called the α-domain which has significant ... The yeast MATα1 is required for the activation of α-specific genes in Saccharomyces cerevisiae and thus confers the α-cell identity of the yeast. MATα1 contains a domain called the α-domain which has significant sequence identity to the HMG-box family of proteins. A multiple sequence alignment of several α-domains and various structurally determined HMG-box domains has revealed that both domains possess very similar structural and functional residues. We found that the basic amino acids of the N-terminal loop, the intercalating hydrophobic residues of the first helix, and the hydrophobic residues required for interactions within the core of the protein are remarkably conserved in α-domains and HMG-box proteins. Our generated molecular models suggest that the first and third helix will be shorter and that the HMG-box core is not an isolated domain. The region beyond the conserved HMG-box motif contains an extended helical region for about 20 - 30 amino acids. Structural models generated by comparative modeling and ab initio modeling reveal that this region will add two or more additional α-helices and will make significant contacts to helix III, II and I of the HMG-box core. We were able to illustrate how the extended α-domain would bind to DNA by merging of the α-domain and the LEF-1/DNA complex. The models we are reporting will be helpful in understanding how MATα1 binds to DNA with its partner MCM1 and activates transcription of α-specific genes. These models will also aid in future biophysical studies of MATα1 including the crystallization and structure determination. 展开更多
关键词 MATα1 MATα2 gene Regulation MATING-TYPE YEAST α-domain Combinatorial Control of Transcription
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Bcl-2 loop domain在Bcl-2蛋白结构中的作用初探
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作者 刘娜 孙志扬 +2 位作者 李杨 宋海燕 孙庆文 《复旦学报(医学版)》 CAS CSCD 北大核心 2006年第5期671-674,共4页
目的探讨B细胞淋巴瘤/白血病基因2(Bcl-2)Loop domain结构域在Bcl-2蛋白结构中的作用。方法应用PCR及Western blot方法检测Bcl-2 Loop domain结构域突变前后Bcl-2蛋白的表达情况。结果Bcl-2 Loop domain结构域突变后Bcl-2蛋白只在沉淀... 目的探讨B细胞淋巴瘤/白血病基因2(Bcl-2)Loop domain结构域在Bcl-2蛋白结构中的作用。方法应用PCR及Western blot方法检测Bcl-2 Loop domain结构域突变前后Bcl-2蛋白的表达情况。结果Bcl-2 Loop domain结构域突变后Bcl-2蛋白只在沉淀中检测到,在上清中未检测到。结论Bcl-2 Loop domain结构域被突变后,Bcl-2蛋白空间结构发生改变,推测Bcl-2 Loop domain结构域在Bcl-2蛋白结构中起到重要作用。 展开更多
关键词 B细胞淋巴瘤/白血病基因2 环状结构域 基因突变
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Pediatric fatty liver disease:Role of ethnicity and genetics 被引量:5
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作者 Pierluigi Marzuillo Emanuele Miraglia del Giudice Nicola Santoro 《World Journal of Gastroenterology》 SCIE CAS 2014年第23期7347-7355,共9页
Non-alcoholic fatty liver disease (NAFLD) comprehends a wide range of conditions, encompassing from fatty liver or steatohepatitis with or without fibrosis, to cirrhosis and its complications. NAFLD has become the mos... Non-alcoholic fatty liver disease (NAFLD) comprehends a wide range of conditions, encompassing from fatty liver or steatohepatitis with or without fibrosis, to cirrhosis and its complications. NAFLD has become the most common form of liver disease in childhood as its prevalence has more than doubled over the past 20 years, paralleling the increased prevalence of childhood obesity. It currently affects between 3% and 11% of the pediatric population reaching the rate of 46% among overweight and obese children and adolescents. The prevalence of hepatic steatosis varies among different ethnic groups. The ethnic group with the highest prevalence is the Hispanic one followed by the Caucasian and the African-American. This evidence suggests that there is a strong genetic background in the predisposition to fatty liver. In fact, since 2008 several common gene variants have been implicated in the pathogenesis of fatty liver disease. The most important is probably the patatin like phospholipase containing domain 3 gene (PNPLA3) discovered by the Hobbs&#x02019; group in 2008. This article reviews the current knowledge regarding the role of ethnicity and genetics in pathogenesis of pediatric fatty liver. 展开更多
关键词 Non alcoholic fatty liver disease ETHNICITY Patatin like phospholipase containing domain 3 gene Obesity Insulin resistance Glucokinase regulatory protein Apolipoprotein C3 gene Farnesyl-diphosphate farnesyltransferase 1
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Genetics of non-alcoholic fatty liver disease: From susceptibility and nutrient interactions to management 被引量:3
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作者 Vishnubhotla Venkata Ravi Kanth Mitnala Sasikala +2 位作者 Mithun Sharma Padaki Nagaraja Rao Duvvuru Nageshwar Reddy 《World Journal of Hepatology》 CAS 2016年第20期827-837,共11页
Genetics plays an important role in determining the susceptibility of an individual to develop a disease. Complex, multi factorial diseases of modern day(diabetes, cardiovascular disease, hypertension and obesity) are... Genetics plays an important role in determining the susceptibility of an individual to develop a disease. Complex, multi factorial diseases of modern day(diabetes, cardiovascular disease, hypertension and obesity) are a result of disparity between the type of food consumed and genes, suggesting that food which does not match the host genes is probably one of the major reasons for developing life style diseases. Non-alcoholic fatty liver is becoming a global epidemic leading to substantial morbidity. While various genotyping approaches such as whole exome sequencing using next generation sequencers and genome wide association studies have identified susceptibility loci for non-alcoholic fatty liver disease(NAFLD) including variants in patatin-like phospholipase domain containing 3 and transmembrane 6 superfamily member 2 genes apart from others; nutrient based studies emphasized on a combination of vitamin D, E and omega-3 fatty acids to manage fatty liver disease. However majority of the studies were conducted independent of each other and very few studies explored the interactions between the genetic susceptibility and nutrient interactions. Identifying such interactions will aid in optimizing the nutrition tailor made to an individual's genetic makeup, thereby aiding in delaying the onset of the disease and its progression. The present topic focuses on studies that identified the genetic susceptibility for NAFLD, nutritional recommendations, and their interactions for better management of NAFLD. 展开更多
关键词 Transmembrane 6 superfamily member 2 gene Patatin-like phospholipase domain containing 3 gene GENOTYPING Nutrient interactions Non-alcoholic fatty liver disease genetic susceptibility
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Chromatin boundary elements organize genomic architecture and developmental gene regulation in Drosophila Hox clusters 被引量:1
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作者 Zhibo Ma Mo Li +5 位作者 Sharmila Roy Kevin J Liu Matthew L Romine Derrick C Lane Sapna K Patel Haini N Cai 《World Journal of Biological Chemistry》 CAS 2016年第3期223-230,共8页
The three-dimensional(3D) organization of the eukaryotic genome is critical for its proper function. Evidence suggests that extensive chromatin loops form the building blocks of the genomic architecture, separating ge... The three-dimensional(3D) organization of the eukaryotic genome is critical for its proper function. Evidence suggests that extensive chromatin loops form the building blocks of the genomic architecture, separating genes and gene clusters into distinct functional domains. These loops are anchored in part by a special type of DNA elements called chromatin boundary elements(CBEs). CBEs were originally found to insulate neighboring genes by blocking influences of transcriptional enhancers or the spread of silent chromatin. However, recent results show that chromatin loops can also play a positive role in gene regulation by looping out intervening DNA and "delivering" remote enhancers to gene promoters. In addition, studies from human and model organisms indicate that the configuration of chromatin loops, many of which are tethered by CBEs, is dynamically regulated during cell differentiation. In particular, a recent work by Li et al has shown that the SF1 boundary, located in the Drosophila Hox cluster, regulates local genes by tethering different subsets of chromatin loops: One subset enclose a neighboring gene ftz, limiting its access by the surrounding Scr enhancers and restrict the spread of repressive histones during early embryogenesis; and the other loops subdivide the Scr regulatory region into independent domains of enhancer accessibility. The enhancer-blocking activity of these CBE elements varies greatly in strength and tissue distribution. Further, tandem pairing of SF1 and SF2 facilitate the bypass of distal enhancers in transgenic flies, providing a mechanism for endogenous enhancers to circumvent genomic interruptions resulting from chromosomal rearrangement. This study demonstrates how a network of chromatin boundaries, centrally organized by SF1, can remodel the 3D genome to facilitate gene regulation during development. 展开更多
关键词 CHROMATIN boundary element Insulator CTCF CHROMATIN loop domains DROSOPHILA HOX genes
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Identification and Characterization of Gene SpDMRT99B and Its Sex-Biased Expression Profile in the Mud Crab,Scylla paramamosain 被引量:1
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作者 ZHANG Yin FANG Shaobin +5 位作者 LIN Fei LI Shengkang ZHENG Huaiping ZHANG Yueling IKHWANUDDIN Mhd MA Hongyu 《Journal of Ocean University of China》 SCIE CAS CSCD 2021年第6期1495-1504,共10页
The doublesex and mab-3 related transcription factor(DMRT)gene family is conserved from invertebrates to humans.The functions of DMRT are mainly involved in sex development and the formation of many tissues and organs... The doublesex and mab-3 related transcription factor(DMRT)gene family is conserved from invertebrates to humans.The functions of DMRT are mainly involved in sex development and the formation of many tissues and organs.In this study,a DM(Doublesex/Mab-3-domain gene was identified in the mud crab Scylla paramamosain,and was named SpDMRT99B because of its many similarities to arthropod DMRT99B and phylogenetically close relationship with arthropod DMRT99B.The cDNA of SpDMRT99B gene is 1249 bp in length,encoding 224 amino acids.From 254 bp to 928 bp there is a conserved DM domain.No transmembrane domain was identified.Through multiple amino acid alignment and phylogenetic tree analysis,the closest gene to SpDMRT99B is Eriocheir sinensis DMRT99B,followed by Macrobrachium rosenbergii and Drosophila melanogaster DMRT99B.The expressions of the gene were characterized in different tissues of female and male crabs during early development period of crab individuals,as well as in different development periods of gonads.The results showed that SpDMRT99B gene is significantly highly expressed in testis than in ovary and other tissues.The expression level of SpDMRT99B in testis at different stages is significantly higher than that in ovary,and it is particularly highly expressed in immature testis.In early developmental stages of larvae,the expressions of SpDMRT99B remain at a low level and reach a peak at zoea stage I when the body segments shape up.It is speculated that SpDMRT99B gene might be involved in the gonadal development process and somitogenesis of S.paramamosain. 展开更多
关键词 Scylla paramamosain DM domain gene SpDMRT99B sex determination/differentiation early development stage
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Complete androgen insensitivity syndrome caused by the c.2678C>T mutation in the androgen receptor gene:A case report 被引量:1
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作者 Ka-Na Wang Qing-Qing Chen +1 位作者 Yi-Lin Zhu Chun-Lin Wang 《World Journal of Clinical Cases》 SCIE 2021年第35期11036-11042,共7页
BACKGROUND Androgen insensitivity syndrome is an X-linked recessive genetic disease caused by mutations in the androgen receptor gene(AR).However,the underlying molecular mechanisms for the majority of AR variants rem... BACKGROUND Androgen insensitivity syndrome is an X-linked recessive genetic disease caused by mutations in the androgen receptor gene(AR).However,the underlying molecular mechanisms for the majority of AR variants remain unclear.In this study,we identified a point variant in three patients with complete androgen insensitivity syndrome(CAIS),summarized the correlation analysis,and performed a literature review.CASE SUMMARY The proband was raised as a girl.In infancy,she was first referred to hospital with a right inguinal hernia.Ultrasonography revealed the absence of a uterus and ovaries,and a testis-like structure located at the inguinal canal.Further diagnostic workup detected a 46,XY karyotype,and fluorescence in situ hybridization analysis showed the presence of the SRY gene.Histological analysis revealed the excised tissue to be testicular.Twelve years later,she was admitted to our hospital with a lack of breast development.Her pubic hair and breasts were Tanner stage I.She had normal female external genitalia.Blood hormone tests showed normal testosterone levels,low estradiol levels,and high gonadotropin levels.Her two siblings underwent similar examinations,and all three had a rare hemizygous missense mutation in AR:c.2678C>T.In vitro functional analyses revealed decreased nuclear translocation in AR-c.2678C>T mutation cells.CONCLUSION This case of CAIS was caused by an AR variant(c.2678C>T).Functional studies showed impaired nuclear translocation ability of the mutant protein. 展开更多
关键词 Androgen insensitivity syndrome 46 XY disorders of sex development VARIANTS Androgen receptor gene Ligand-binding domain Case report
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Cloning of WWOX Gene and Its Growth-inhibiting Effects on Ovarian Cancer Cells 被引量:2
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作者 熊宙芳 胡沙 王泽华 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2010年第3期365-369,共5页
The growth-inhibiting and apoptosis-inducing effects of WW domain-containing oxidoreductase(WWOX) gene on ovarian cancer cell line A2780 were investigated.The full length cDNA of human WWOX gene was amplified from nor... The growth-inhibiting and apoptosis-inducing effects of WW domain-containing oxidoreductase(WWOX) gene on ovarian cancer cell line A2780 were investigated.The full length cDNA of human WWOX gene was amplified from normal human ovary tissues.The correct cDNA of full length WWOX was subcloned into eukaryocytic expression vector pCMV.After introduction of WWOX gene into cancer cells with liposome,the WWOX mRNA and protein level in the cancer cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and immunoblotting.The growth activities of cancer cells were detected by Trypan blue staining.The clone formation assay in soft agar was employed to observe the proliferation of the cancer cells.Apoptosis was examined by DNA ladder and acridine orange-ethidium bromide fluorescent staining.The results showed that 72 h after WWOX gene transfection,the WWOX expression was increased significantly(P<0.01).The growth of ovarian cancer cells was decreased by 16.41% to 38.49%(P<0.01).The clone formation abilities were reduced(P<0.01).Some cancer cells presented the characteristic morphological changes of apoptosis with obvious ladder bands on electrophoresis.The apoptosis rate was(20.7±6.0)%(P<0.01).It was concluded that over-expression of WWOX gene could induce apoptosis and inhibit the growth of ovarian cancer cells,which might be potentially useful in the gene therapy of ovarian cancers. 展开更多
关键词 gene expression ovarian neoplasms WW domain-containing oxidoreductase APOPTOSIS
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Expression of Core Domain of Porcine Zona Pellucida 3β in E.coli
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作者 Qiu-ling XIE Xiao-jia CHEN +5 位作者 Wei-jie ZHU Ling ZHANG Wan-xiang XU An HONG Jing LI Si-hong GAO 《Journal of Reproduction and Contraception》 CAS 2005年第2期67-72,共6页
To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained ... To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained by PCR and then was cloned into pET-3c vector. After being identified, recon was transformed into E.coli BL21 (DE3) pLysS and then induced by IPTG. Results The recombinant cZP3β was expressed in E. coli up to 15% of total cellular proteins, and was made sure by Western blot analysis. Conclusion The research on expression of core domain of pZP3β could benefit to further investigation of its immunogenicity and the development of antigen preparation. 展开更多
关键词 porcine zona pellucida core domain E. coli gene expression porcine zona pellucida core domain E. coli gene expression
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A Novel Bacillus thuringiensis Cry57 Protein Domain Swap Influence on Insecticidal Activity
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作者 Shi Chen-chen Zhou Guo-wang +2 位作者 Liu Zhi-yang Gao Ji-guo Li Hai-tao 《Journal of Northeast Agricultural University(English Edition)》 CAS 2018年第3期44-52,共9页
Bacillus thuringiensis (Bt) is widely used in insecticides. Bt is a gram positive sporulation bacterium belonging to Bacillaceae family. It produces different insecticidal proteins like Cry toxin, Vip toxin, β-exot... Bacillus thuringiensis (Bt) is widely used in insecticides. Bt is a gram positive sporulation bacterium belonging to Bacillaceae family. It produces different insecticidal proteins like Cry toxin, Vip toxin, β-exotoxin and chitinase, etc. Bt insecticidal crystal proteins (ICPs) are not homologous to other known Vip protein and then act against lepidopteran, dipteran, coleopteran and nematodeslarvae via a unique process. In this experiment, modern high-throughput sequencing technique and sequencing were used and the whole genome sequence of BtLTS290 was obtained. The results compared to the database of GenBank showed that there was a cry57 gene in the genome sequence of BtLTS290. A novel cry57 gene was cloned and named cry57Ab1 (accession number is KF638650) by International Nomenclature Committee of Bt Endotoxin. cry57Ab1 gene could be expressed with the molecular weight of 90 ku. Cry57Ab1 protein had no obvious activity against Spodoptera exigua and Helicoverpa armigera. And Cry57Ab1 protein had a slight insecticidal activity against Ostrinia furnacalis and Plutella xylostella. Furthermore, the domain Ⅱof Cry57Ab1 and Cry1Bb were exchanged by overlapping extension PCR. SDS-PAGE showed that the molecular weight of Cry57Ab/1Bb/57Ab was about 90 ku. The insecticidal activity of Cry57Ab1 protein and Cry57Ab/1Bb/57Ab recombinant protein were determined. The results showed that the insecticidal activity of the recombinant protein to Spodoptera exigua and Helicoverpa armigera was very low, and the corrected mortality was less than 10%. The insecticidal activities against Ostrinia furnacalis and Plutella xylostella were reduced. The corrected mortality of Ostrinia furnacalis was 4.4%, and the corrected mortality rate for Plutella xylostella was 6.7%. Domain Ⅱof cry toxin played a key role on affecting host specifcity. 展开更多
关键词 Bacillus thuringiensis cry57Ab1 gene domain exchange BIOASSAY
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Effect of Antisense MBD1 Gene Eukaryotic Expression Plasmid on Expression of MBD1 Gene in Human Biliary Tract Carcinoma Cells
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作者 左石 邹声泉 +4 位作者 罗剑 郭伟 徐立宁 董泾青 刘民锋 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2005年第6期658-661,共4页
Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumori... Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912±0.022 to 0. 215±0. 017, and the protein level of MBD1 gene also decreased from (80.19±5.05) %to (35.11±4.05) %. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression plasmid transfection group (P〈0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma. 展开更多
关键词 methyl-CpG binding domain protein 1 antisense RNA TRANSFECTION gene expression biliary tract carcinoma
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Mutational Analyses of a Fork Head Associated Domain Protein, DAWDLE, in <i>Arabidopsis thaliana</i>
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作者 Lakshmi Ayiloor Narayanan Dipaloke Mukherjee +2 位作者 Shuxin Zhang Bin Yu David Chevalier 《American Journal of Plant Sciences》 2014年第18期2811-2822,共12页
DAWDLE (DDL) gene encodes a protein that contains an N-terminal arginine-rich domain and a C-terminal Fork Head Associated (FHA) domain in Arabidopsis thaliana. DDL protein is believed to function in microRNA biogenes... DAWDLE (DDL) gene encodes a protein that contains an N-terminal arginine-rich domain and a C-terminal Fork Head Associated (FHA) domain in Arabidopsis thaliana. DDL protein is believed to function in microRNA biogenesis by mediating the recruitment of pri-microRNA to DICER-LIKE 1 and also stabilizing the microRNA. The aim of this study was to conduct a structure-function analysis to identify the regions in DDL that are of functional significance. Targeted Induced Local Lesions in Genome screen was performed in the Columbia erecta-105 background of Arabidopsis resulting in the identification of eight point mutations spanning DDL. The mutants were characterized by phenotypic and molecular analyses based on the prior knowledge on ddl knockout mutants. Height of the plant, hypocotyl and root length, and fertility were measured for phenotypic characterization, and microRNA172 levels were measured to assess the mutation effect at the molecular level. Phenotypic and molecular analyses of the mutants revealed effects resulting in ddl phenotypes of varying degrees in different organs and each mutant displayed at least one phenotype studied. Reduction in fertility and increase in stem length were two phenotypes that most of the mutants consistently displayed. Identification and characterization of several key residues in the arginine rich region and FHA domain will serve as an important tool for elucidation of DDL signaling pathway. 展开更多
关键词 microRNA DAWDLE FHA domain gene Mapping ARABIDOPSIS
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Newly identified genetic variant rs2294693 in UNC5CL gene is associated with decreased risk of esophageal carcinoma in the J&K Population–India
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作者 RUCHI SHAH SONALI VERMA +7 位作者 AMRITA BHAT GH RASOOL BHAT VARUN SHARMA INDU SHARMA HEMENDER SINGH SANDEEP KAUL EKTA RAI SWARKAR SHARMA 《BIOCELL》 SCIE 2021年第3期665-670,共6页
Esophageal cancer is the second most common type of cancer after lung carcinoma in the state of Jammu and Kashmir(J&K).The understanding of genetics in Esophageal cancer development is poor in the state.Genome wid... Esophageal cancer is the second most common type of cancer after lung carcinoma in the state of Jammu and Kashmir(J&K).The understanding of genetics in Esophageal cancer development is poor in the state.Genome wide association studies(GWAS)has proved to be unsurpassed tool in identification of new loci associated with different cancers.GWAS in Chinese population has identified SNP rs2294693 present in UNC5CL(UNC-5 Family C-Terminal like)to be associated with non-cardia gastric cancer.We performed a case control association study and genotyped the SNP rs2294693 using Taqman allele discrimination assay in 566 individuals(166 esophageal cancer patients and 400 controls)belonging to the J&K population.A statistically significant protective association with allelic odds ratio of 0.73(0.56–0.94 at 95%CI)and p value=0.016 was observed.This is the first study in relation to esophageal cancer in the Jammu and Kashmir population,so far it has been studied in association with gastric carcinoma in the Chinese population only.The results indicate that the polymorphism rs2294693 is associated with esophageal cancer susceptibility and the mutant(T)allele might be a protective factor for esophageal cancer among Jammu and Kashmir population.Further the functional characterization of the variation is also warranted. 展开更多
关键词 Esophageal cancer Single nucleotide polymorphism Unc-5 Family c-terminal(UNC5CL) Candidate gene
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Characterization of a TaJ Gene from Wheat
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作者 LI Guo-liang CHANG Hui ZHOU Ren-gang 《Agricultural Sciences in China》 CAS CSCD 2007年第9期1043-1050,共8页
A novel J-domain protein gene was cloned from wheat (Triticum aestivum L.) using RT-PCR technology and named as TaJ. The J-domain protein is defined by the presence of a J-domain. The cDNA of T. aestivum gene, TaJ ... A novel J-domain protein gene was cloned from wheat (Triticum aestivum L.) using RT-PCR technology and named as TaJ. The J-domain protein is defined by the presence of a J-domain. The cDNA of T. aestivum gene, TaJ (GenBank accession number: DQ789026), was 1263 bp and contained a complete open reading frame (ORF) encoding a J-domain protein of 420 amino acid residues. The predicted amino acid sequence of TaJ possesses three functionally essential domains: the Nterminal J-domain which includes the highly conserved HPD tripeptide, an adjacent domain that is rich in glycine and phenylalanine residues (G/F) and a Cysteine-rich zinc-finger domain with four repeats of CxxCxGxG that is important for protein interactions. The C-terminal of TaJ was -CAQQ, a farnesylation motif. The full-length deduced amino acid sequence of TaJ is highly homologous to J-domain proteins from various plant species. Southern blot analysis indicated that a single copy of TaJ existed in wheat genome. The expression pattern of TaJ performed by real-time PCR demonstrated that heat shock (HS) at 37℃ induced the expression of TaJ rapidly and strongly, but the response of the TaJ gene to cold stress was much slower than that to HS. Tissue-specific expression analysis showed that the expression level of TaJ gene was much higher in leaves than that in roots. 展开更多
关键词 WHEAT J-domain protein CLONING gene expression heat shock
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