High temperature stress is one of the major environmental factors that affect the growth and development of plants. Although WRKY transcription factors play a critical role in stress responses, there are few studies o...High temperature stress is one of the major environmental factors that affect the growth and development of plants. Although WRKY transcription factors play a critical role in stress responses, there are few studies on the regulation of heat stress by WRKY transcription factors,especially in tomato. Here, we identified a group I WRKY transcription factor, SlWRKY3, involved in thermotolerance in tomato. First, SlWRKY3 was induced and upregulated under heat stress. Accordingly, overexpression of SlWRKY3 led to an increase, whereas knock-out of SlWRKY3 resulted in decreased tolerance to heat stress. Overexpression of SlWRKY3 accumulated less reactive oxygen species(ROS), whereas knock-out of SlWRKY3 accumulated more ROS under heat stress. This indicated that SlWRKY3 positively regulates heat stress in tomato. In addition,SlWRKY3 activated the expression of a range of abiotic stress-responsive genes involved in ROS scavenging, such as a SlGRXS1 gene cluster.Further analysis showed that SlWRKY3 can bind to the promoters of the SlGRXS1 gene cluster and activate their expression. Collectively, these results imply that SlWRKY3 is a positive regulator of thermotolerance through direct binding to the promoters of the SlGRXS1 gene cluster and activating their expression and ROS scavenging.展开更多
This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and im...This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and immunohis to chemically detected to evaluate the correlation of EMP-1 expression to the clinical features of NSCLC. Recombinant adenovirus was constructed to over-express EMP-1 and then infect PC9 cells. Cell proliferation was measured by Ki67 staining. Western blotting was performed to examine the effect of EMP-1 on the PI3K/AKT signaling. Moreover, tumor xeno-grafts were established by subcutaneous injection of PC9 cell suspension (about 5×107/mL in 100 μL of PBS) into the right hind limbs of athymic nude mice. The results showed EMP-1 was significantly up-regulated in NSCLC patients as compared with those with benign lung diseases. Over-expression of EMP-1 promoted proliferation of PC9 cells, which coincided with the activation of the PI3K/AKT pathway. EMP-1 promoted the growth of xenografts of PC9 cells in athymic nude mice. It was concluded that EMP-1 expression may contribute to the development and progress of NSCLC by activating PI3K/AKT pathway.展开更多
Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321&#...Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.展开更多
BACKGROUND Pancreatic cancer is a highly invasive malignant tumor. Expression levels of the autophagy-related protein microtubule-associated protein 1 A/1 B-light chain 3(LC3) and perineural invasion(PNI) are closely ...BACKGROUND Pancreatic cancer is a highly invasive malignant tumor. Expression levels of the autophagy-related protein microtubule-associated protein 1 A/1 B-light chain 3(LC3) and perineural invasion(PNI) are closely related to its occurrence and development. Our previous results showed that the high expression of LC3 was positively correlated with PNI in the patients with pancreatic cancer. In this study, we further searched for differential genes involved in autophagy of pancreatic cancer by gene expression profiling and analyzed their biological functions in pancreatic cancer, which provides a theoretical basis for elucidating the pathophysiological mechanism of autophagy in pancreatic cancer and PNI.AIM To identify differentially expressed genes involved in pancreatic cancer autophagy and explore the pathogenesis at the molecular level.METHODS Two sets of gene expression profiles of pancreatic cancer/normal tissue(GSE16515 and GSE15471) were collected from the Gene Expression Omnibus.Significance analysis of microarrays algorithm was used to screen differentially expressed genes related to pancreatic cancer. Gene Ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis were used to analyze the functional enrichment of the differentially expressed genes. Protein interaction data containing only differentially expressed genes was downloaded from String database and screened. Module mining was carried out by Cytoscape software and ClusterOne plug-in. The interaction relationship between the modules was analyzed and the pivot nodes between the functional modules were determined according to the information of the functional modules and the data of reliable protein interaction network.RESULTS Based on the above two data sets of pancreatic tissue total gene expression, 6098 and 12928 differentially expressed genes were obtained by analysis of genes with higher phenotypic correlation. After extracting the intersection of the two differential gene sets, 4870 genes were determined. GO analysis showed that 14 significant functional items including negative regulation of protein ubiquitination were closely related to autophagy. A total of 986 differentially expressed genes were enriched in these functional items. After eliminating the autophagy related genes of human cancer cells which had been defined, 347 differentially expressed genes were obtained. KEGG pathway analysis showed that the pathways hsa04144 and hsa04020 were related to autophagy. In addition,65 clustering modules were screened after the protein interaction network was constructed based on String database, and module 32 contains the LC3 gene,which interacts with multiple autophagy-related genes. Moreover, ubiquitin C acts as a pivot node in functional modules to connect multiple modules related to pancreatic cancer and autophagy.CONCLUSION Three hundred and forty-seven genes associated with autophagy in human pancreatic cancer were concentrated, and a key gene ubiquitin C which is closely related to the occurrence of PNI was determined, suggesting that LC3 may influence the PNI and prognosis of pancreatic cancer through ubiquitin C.展开更多
The objective of this study was to determine the genotype effects of the bovine insulin-like growth factor Ⅰ （IGF-Ⅰ） and its binding protein 3 （IGFBP3） genes on growth and development traits in beef cows, includ...The objective of this study was to determine the genotype effects of the bovine insulin-like growth factor Ⅰ （IGF-Ⅰ） and its binding protein 3 （IGFBP3） genes on growth and development traits in beef cows, including 130 Chinese Simmental, 42 Nanyang, and 47 Luxi Yellow cattle. Sequence variations in the bovine IGF-Ⅰ and IGFBP3 genes were investigated by single strand conformation polymorphism （SSCP）. SSCPs were detected in 6 fragments, which is the 5＇-flanking region, the 2nd exon, the 5th exon, and the 5th intron of the IGF-1 gene, and the 2nd exon, the 3rd exon of the 1GFBP3 gene. Two polymorphisms, an A-to-G transition in the 2rid exon of the IGF-Ⅰ gene and a T-to-C transition in the 2rid exon of IGFBP3 gene were detected in 3 breeds. The allele frequencies of 2 polymorphisms were 0.0411 （A）, 0.9589 （B）, and 0.7237 （A）, 0.2763 （B）, respectively. These 2 loci were analyzed to associate with body weight, height at withers, body length, heart girth, rump width, and beef production index （BPI） at 0, 6, 12, 24, and 36-month old. The IGFBP3 locus was shown to be associated with rump width, heart girth at 24-month and 36-month. Animals with BB genotype had higher rump width （24.86 ± 0.47） cm at 24-month and （27.50 ± 0.63） em at 36-month. The heart girth was highest for the individuals with BB genotype （171.33 ± 1.84） cm and higher than those with AB genotype （166.68 ± 1.13） cm （P〈 0.05） at 36-month.展开更多
AIM： To identify the disease-causing mutation responsible for the presence of congenital cataract in a Chinese family. METHODS： The study recruited a four-generation Chinese pedigree affected by autosomal dominant c...AIM： To identify the disease-causing mutation responsible for the presence of congenital cataract in a Chinese family. METHODS： The study recruited a four-generation Chinese pedigree affected by autosomal dominant congenital cataract （ADCC）. Family history and the history of cataract extraction were recorded. Blood samples were collected from individuals for DNA extraction. Direct sequencing of congenital cataract-associated genes was performed. Single-strand conformational polymorphism and bioinformatic analysis were conducted to further study the mutation. RESULTS： Direct sequencing revealed a novel splice site mutation of c.30-2 A〉G in the CRYBA3/A1 gene. The mutation co-segregated within all affected individuals in the family and was not found in unaffected members or 100 unrelated normal controls. These results were further confirmed by single-strand conformational polymorphism and bioinformatic analysis using the Human Splicing Finder and MaxEnt online software and Annovar computer software. CONCLUSION： c,30-2 A〉G mutation of CRYBA3/A1 gene is a novel mutation and broadens the genetic spectrum of ADCC, KEYWORDS： splice site mutation; congenital cataract; CRYBA3/A1 gene展开更多
Hybrid rice Fanyou 7206（FY7206）, derived from the cross between a sterile line Fanyuan A and a restorer line Fuhui 7206, was bred by the Rice Research Institute, Fujian Academy of Agricultural Sciences, China. FY720...Hybrid rice Fanyou 7206（FY7206）, derived from the cross between a sterile line Fanyuan A and a restorer line Fuhui 7206, was bred by the Rice Research Institute, Fujian Academy of Agricultural Sciences, China. FY7206 was characterized by moderate blast resistance, cold tolerance, as well as wide adaptability, and high yields. The blast resistance results indicated that the frequencies of blast races in race B, race C and the total resistance frequency for FY7206 were 95.5%, 100.0% and 97.2%, respectively. The disease resistance results showed that the leaf blast grade for FY7206 was level 1 and panicle blast was level 5. The indoor spray results indicated that FY7206 was resistant to 11 isolates of Magnorpathe oryzae. The blast resistance of FY7206 might be derived from the high expression of blast resistance gene Pid3. The results for simulated cold resistance in an artificial climate chamber indicated that the cold tolerance for FY7206 was moderate at the booting and flowering stages. The cold tolerance results also indicated that FY7206 could be tolerant to temperatures as low as 10 °C at the seedling stage. The q RT-PCR results showed that the expression of cold tolerance gene Ctb1 in FY7206 was relatively high. These results suggested that FY7206 is a hybrid indica rice variety with good comprehensive characteristics, including blast resistance and cold tolerance.展开更多
Bipolaris oryzae is the causal agent of brown spot disease in rice and produces the dark pigment melanin. We isolated and characterized T4HR1 gene encoding 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN) reductase, whic...Bipolaris oryzae is the causal agent of brown spot disease in rice and produces the dark pigment melanin. We isolated and characterized T4HR1 gene encoding 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN) reductase, which converted 1,3,6,8-THN to scytalone in the melanin biosynthesis from B. oryzae. A sequence analysis showed that the T4HR1 gene encoded a putative protein of 268 amino acids showing 50% - 99% sequence identity to other fungal 1,3,6,8-THN reductases. Targeted disruption of the T4HR1 gene showed a different phenotype of mycelial color due to an accumulation of shunt products compared to those of wild-type on PDA plates using tricyclazole as a melanin biosynthesis inhibitor. A quantitative real-time PCR analysis showed that the expression of T4HR1 transcripts was enhanced by near-ultraviolet (NUV) irradiation and regulated by transcriptional factor BMR1, similar to three other melanin biosynthesis genes (polyketide synthase gene [PKS1], scytalone dehydratase gene [SCD1], and 1,3,8-THN reductase gene [THR1]) in the melanin biosynthesis of B. oryzae. These results suggested that common transcriptional mechanisms could regulate the enhanced gene expression of these melanin biosynthesis genes by NUV irradiation in B. oryzae.展开更多
In order to reveal which role the callose played in R. rugosa pollination incompatibility, the full-length cDNA sequence of β-1,3-glucanase gene was cloned for the first time from the stylus of Rosa rugosa “Tanghong...In order to reveal which role the callose played in R. rugosa pollination incompatibility, the full-length cDNA sequence of β-1,3-glucanase gene was cloned for the first time from the stylus of Rosa rugosa “Tanghong” with RT-PCR and RACE methods and named as RrGlu. The full-length cDNA is 1380 bp with an open reading frame of 1041 bp, encoding 346 amino acids. The derived protein has a molecular weight of 37.85 kD, a calculated pI of 9.12, a pfam00332 conserved domain at position 36 - 345, and belongs to glycosyl hydrolase family 17. The derived protein is a hydrophilic protein secreted into the vacuole. There is a signal peptide cleavage site at position 34 - 35, a transmembrane domain at position 13 - 32, six Ser phosphorylation sites, three Thr phosphorylation sites, three Tyr phosphorylation sites, one N-glycosylation site, and five O-glycosylation sites. There are 31.50% α-helixes, 30.92% random coil, 25.14% extended peptide chain, and 12.43% β-corner structure. This protein and the Glu protein from eight other species, including Prunus persica, share a sequence homology of greater than 72%;all of the proteins contain a pfam00332 conserved domain and a β-1,3-glucanase active center sequence (LIVM)-X-(LIVMFYW)3-(STAG)-E-(ST)-G-W-P-(ST)-X-G. Furthermore, their phylogenetic relationships are consistent with their traditional classifications. These results were meaningful to reveal the molecular mechanism of R. rugosa pollination incompatibility and improve the theory and techniques of breeding ornamental R. rugosa.展开更多
HIV-1 infection requires the expression of CD4＋ molecules in colligation with C-C chemokine receptor type 5 （CCR5） and C-X-C chemokine receptor type 4 （CXCR4） as the major coreceptors. The role of SNP in 3＇ untr...HIV-1 infection requires the expression of CD4＋ molecules in colligation with C-C chemokine receptor type 5 （CCR5） and C-X-C chemokine receptor type 4 （CXCR4） as the major coreceptors. The role of SNP in 3＇ untranslated region ofSDF-1 （SDF1-3 ＇A） and low copy number （CN） of the CCL3L1 gene is reported to confer increased resistance to HIV-1 infection. The aim of the present study was to analyze the combinatorial effect of both the variations in protection towards HIV-1 infection in Indian population. The combinatorial effect of genetic variation in terms of SNP in SDF-1 gene and CCL3L1 CN was investigated in 105 healthy individuals and 78 HIV-I patients. Genotyping of SDF-1 was performed by RFLP-PCR and CCL3L1 by real-time PCR using TaqMan chemistry. The genotype frequency distribution of SDF-1 was found to be （SDF-1/SDF-I： 65.4%, SDF-1/SDF1-3＇A： 29.5% and SDFI-3＇A/SDF1-3＇A- 5.1%） in HIV patients as compared to （SDF-1/SDF-I： 64.8%, SDF-1/SDF1-3＇A： 30.5% and SDF1-3 ＇A/SDF1-3 ＇A： 4.7%） in healthy individuals, whereas a range of 1 to 6 copies per diploid genome was observed for CCL3L1 gene.展开更多
基金supported by grants from the National Key Research&Development Plan,China (Grant Nos.2021YFD1200201,2022YFD1200502)National Natural Science Foundation of China(31972426,31991182)+3 种基金Key Project of Hubei Hongshan Laboratory(Grant No.2021hszd007)Wuhan Major Project of Key Technologies in Biological Breeding (Grant No.2022021302024852)Fundamental Research Funds for the Central Universities,China (Grant No.2662022YLPY001)International Cooperation Promotion Plan of Shihezi University (Grant No.GJHZ202104)。
文摘High temperature stress is one of the major environmental factors that affect the growth and development of plants. Although WRKY transcription factors play a critical role in stress responses, there are few studies on the regulation of heat stress by WRKY transcription factors,especially in tomato. Here, we identified a group I WRKY transcription factor, SlWRKY3, involved in thermotolerance in tomato. First, SlWRKY3 was induced and upregulated under heat stress. Accordingly, overexpression of SlWRKY3 led to an increase, whereas knock-out of SlWRKY3 resulted in decreased tolerance to heat stress. Overexpression of SlWRKY3 accumulated less reactive oxygen species(ROS), whereas knock-out of SlWRKY3 accumulated more ROS under heat stress. This indicated that SlWRKY3 positively regulates heat stress in tomato. In addition,SlWRKY3 activated the expression of a range of abiotic stress-responsive genes involved in ROS scavenging, such as a SlGRXS1 gene cluster.Further analysis showed that SlWRKY3 can bind to the promoters of the SlGRXS1 gene cluster and activate their expression. Collectively, these results imply that SlWRKY3 is a positive regulator of thermotolerance through direct binding to the promoters of the SlGRXS1 gene cluster and activating their expression and ROS scavenging.
基金supported by grants from the National Natural Science Foundation of China (Nos.81072431,30872472,30973496 and 30800569)the Innovative Foundation of Huazhong University of Science and Technology (No.2010MS027)+1 种基金the Foundation of "973" Program (No.2009CB521802)by Special Fund for Central University Basic Scientific Research (Nos.2011JC062,2011JC063)
文摘This study examined the role of EMP-1 in tumorigenesis of non-small cell lung carcinoma (NSCLC) and the possible mechanism. Specimens were collected from 28 patients with benign lung diseases and 28 with NSCLC, and immunohis to chemically detected to evaluate the correlation of EMP-1 expression to the clinical features of NSCLC. Recombinant adenovirus was constructed to over-express EMP-1 and then infect PC9 cells. Cell proliferation was measured by Ki67 staining. Western blotting was performed to examine the effect of EMP-1 on the PI3K/AKT signaling. Moreover, tumor xeno-grafts were established by subcutaneous injection of PC9 cell suspension (about 5×107/mL in 100 μL of PBS) into the right hind limbs of athymic nude mice. The results showed EMP-1 was significantly up-regulated in NSCLC patients as compared with those with benign lung diseases. Over-expression of EMP-1 promoted proliferation of PC9 cells, which coincided with the activation of the PI3K/AKT pathway. EMP-1 promoted the growth of xenografts of PC9 cells in athymic nude mice. It was concluded that EMP-1 expression may contribute to the development and progress of NSCLC by activating PI3K/AKT pathway.
文摘Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.
基金Supported by the National Natural Science Foundation of China,No.U1504815 and No.U1504808
文摘BACKGROUND Pancreatic cancer is a highly invasive malignant tumor. Expression levels of the autophagy-related protein microtubule-associated protein 1 A/1 B-light chain 3(LC3) and perineural invasion(PNI) are closely related to its occurrence and development. Our previous results showed that the high expression of LC3 was positively correlated with PNI in the patients with pancreatic cancer. In this study, we further searched for differential genes involved in autophagy of pancreatic cancer by gene expression profiling and analyzed their biological functions in pancreatic cancer, which provides a theoretical basis for elucidating the pathophysiological mechanism of autophagy in pancreatic cancer and PNI.AIM To identify differentially expressed genes involved in pancreatic cancer autophagy and explore the pathogenesis at the molecular level.METHODS Two sets of gene expression profiles of pancreatic cancer/normal tissue(GSE16515 and GSE15471) were collected from the Gene Expression Omnibus.Significance analysis of microarrays algorithm was used to screen differentially expressed genes related to pancreatic cancer. Gene Ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis were used to analyze the functional enrichment of the differentially expressed genes. Protein interaction data containing only differentially expressed genes was downloaded from String database and screened. Module mining was carried out by Cytoscape software and ClusterOne plug-in. The interaction relationship between the modules was analyzed and the pivot nodes between the functional modules were determined according to the information of the functional modules and the data of reliable protein interaction network.RESULTS Based on the above two data sets of pancreatic tissue total gene expression, 6098 and 12928 differentially expressed genes were obtained by analysis of genes with higher phenotypic correlation. After extracting the intersection of the two differential gene sets, 4870 genes were determined. GO analysis showed that 14 significant functional items including negative regulation of protein ubiquitination were closely related to autophagy. A total of 986 differentially expressed genes were enriched in these functional items. After eliminating the autophagy related genes of human cancer cells which had been defined, 347 differentially expressed genes were obtained. KEGG pathway analysis showed that the pathways hsa04144 and hsa04020 were related to autophagy. In addition,65 clustering modules were screened after the protein interaction network was constructed based on String database, and module 32 contains the LC3 gene,which interacts with multiple autophagy-related genes. Moreover, ubiquitin C acts as a pivot node in functional modules to connect multiple modules related to pancreatic cancer and autophagy.CONCLUSION Three hundred and forty-seven genes associated with autophagy in human pancreatic cancer were concentrated, and a key gene ubiquitin C which is closely related to the occurrence of PNI was determined, suggesting that LC3 may influence the PNI and prognosis of pancreatic cancer through ubiquitin C.
基金funded by the National Key Technology R&D Program of the China (2006BAD01A10,2006BAD14B07, 2006BAD04A16)the National High Technology Research and Development Program of China (2006AA10Z197)
文摘The objective of this study was to determine the genotype effects of the bovine insulin-like growth factor Ⅰ （IGF-Ⅰ） and its binding protein 3 （IGFBP3） genes on growth and development traits in beef cows, including 130 Chinese Simmental, 42 Nanyang, and 47 Luxi Yellow cattle. Sequence variations in the bovine IGF-Ⅰ and IGFBP3 genes were investigated by single strand conformation polymorphism （SSCP）. SSCPs were detected in 6 fragments, which is the 5＇-flanking region, the 2nd exon, the 5th exon, and the 5th intron of the IGF-1 gene, and the 2nd exon, the 3rd exon of the 1GFBP3 gene. Two polymorphisms, an A-to-G transition in the 2rid exon of the IGF-Ⅰ gene and a T-to-C transition in the 2rid exon of IGFBP3 gene were detected in 3 breeds. The allele frequencies of 2 polymorphisms were 0.0411 （A）, 0.9589 （B）, and 0.7237 （A）, 0.2763 （B）, respectively. These 2 loci were analyzed to associate with body weight, height at withers, body length, heart girth, rump width, and beef production index （BPI） at 0, 6, 12, 24, and 36-month old. The IGFBP3 locus was shown to be associated with rump width, heart girth at 24-month and 36-month. Animals with BB genotype had higher rump width （24.86 ± 0.47） cm at 24-month and （27.50 ± 0.63） em at 36-month. The heart girth was highest for the individuals with BB genotype （171.33 ± 1.84） cm and higher than those with AB genotype （166.68 ± 1.13） cm （P〈 0.05） at 36-month.
基金Supported by Key Program of National Natural Science Foundation of China(No.81130018)National NaturalScience Foundation of China(No.81371001+6 种基金No.81570822No.81428005No.81470612)Zhejiang Province Key Research and Development Program(No.2015C03042)Zhejiang Key Lab Fund of China(No.2011E10006)Zhejiang Provincial Natural Science Foundation of China(No.LY14H120002)Zhejiang Province Traditional Chinese medicine Fund project(No.2013ZA082)
文摘AIM： To identify the disease-causing mutation responsible for the presence of congenital cataract in a Chinese family. METHODS： The study recruited a four-generation Chinese pedigree affected by autosomal dominant congenital cataract （ADCC）. Family history and the history of cataract extraction were recorded. Blood samples were collected from individuals for DNA extraction. Direct sequencing of congenital cataract-associated genes was performed. Single-strand conformational polymorphism and bioinformatic analysis were conducted to further study the mutation. RESULTS： Direct sequencing revealed a novel splice site mutation of c.30-2 A〉G in the CRYBA3/A1 gene. The mutation co-segregated within all affected individuals in the family and was not found in unaffected members or 100 unrelated normal controls. These results were further confirmed by single-strand conformational polymorphism and bioinformatic analysis using the Human Splicing Finder and MaxEnt online software and Annovar computer software. CONCLUSION： c,30-2 A〉G mutation of CRYBA3/A1 gene is a novel mutation and broadens the genetic spectrum of ADCC, KEYWORDS： splice site mutation; congenital cataract; CRYBA3/A1 gene
基金supported by grants from the National Program on the Development of Basic Research of China (Grant No. 2013CBA01405-7)the High-Tech Research and Development Program of China (863 Program) (Grant Nos. 2014AA10A603 and 2014AA10A604)the Special Foundation of Non-Profit Research Institutes of Fujian Province, China (Grant No. 2014R1021-15)
文摘Hybrid rice Fanyou 7206（FY7206）, derived from the cross between a sterile line Fanyuan A and a restorer line Fuhui 7206, was bred by the Rice Research Institute, Fujian Academy of Agricultural Sciences, China. FY7206 was characterized by moderate blast resistance, cold tolerance, as well as wide adaptability, and high yields. The blast resistance results indicated that the frequencies of blast races in race B, race C and the total resistance frequency for FY7206 were 95.5%, 100.0% and 97.2%, respectively. The disease resistance results showed that the leaf blast grade for FY7206 was level 1 and panicle blast was level 5. The indoor spray results indicated that FY7206 was resistant to 11 isolates of Magnorpathe oryzae. The blast resistance of FY7206 might be derived from the high expression of blast resistance gene Pid3. The results for simulated cold resistance in an artificial climate chamber indicated that the cold tolerance for FY7206 was moderate at the booting and flowering stages. The cold tolerance results also indicated that FY7206 could be tolerant to temperatures as low as 10 °C at the seedling stage. The q RT-PCR results showed that the expression of cold tolerance gene Ctb1 in FY7206 was relatively high. These results suggested that FY7206 is a hybrid indica rice variety with good comprehensive characteristics, including blast resistance and cold tolerance.
文摘Bipolaris oryzae is the causal agent of brown spot disease in rice and produces the dark pigment melanin. We isolated and characterized T4HR1 gene encoding 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN) reductase, which converted 1,3,6,8-THN to scytalone in the melanin biosynthesis from B. oryzae. A sequence analysis showed that the T4HR1 gene encoded a putative protein of 268 amino acids showing 50% - 99% sequence identity to other fungal 1,3,6,8-THN reductases. Targeted disruption of the T4HR1 gene showed a different phenotype of mycelial color due to an accumulation of shunt products compared to those of wild-type on PDA plates using tricyclazole as a melanin biosynthesis inhibitor. A quantitative real-time PCR analysis showed that the expression of T4HR1 transcripts was enhanced by near-ultraviolet (NUV) irradiation and regulated by transcriptional factor BMR1, similar to three other melanin biosynthesis genes (polyketide synthase gene [PKS1], scytalone dehydratase gene [SCD1], and 1,3,8-THN reductase gene [THR1]) in the melanin biosynthesis of B. oryzae. These results suggested that common transcriptional mechanisms could regulate the enhanced gene expression of these melanin biosynthesis genes by NUV irradiation in B. oryzae.
文摘In order to reveal which role the callose played in R. rugosa pollination incompatibility, the full-length cDNA sequence of β-1,3-glucanase gene was cloned for the first time from the stylus of Rosa rugosa “Tanghong” with RT-PCR and RACE methods and named as RrGlu. The full-length cDNA is 1380 bp with an open reading frame of 1041 bp, encoding 346 amino acids. The derived protein has a molecular weight of 37.85 kD, a calculated pI of 9.12, a pfam00332 conserved domain at position 36 - 345, and belongs to glycosyl hydrolase family 17. The derived protein is a hydrophilic protein secreted into the vacuole. There is a signal peptide cleavage site at position 34 - 35, a transmembrane domain at position 13 - 32, six Ser phosphorylation sites, three Thr phosphorylation sites, three Tyr phosphorylation sites, one N-glycosylation site, and five O-glycosylation sites. There are 31.50% α-helixes, 30.92% random coil, 25.14% extended peptide chain, and 12.43% β-corner structure. This protein and the Glu protein from eight other species, including Prunus persica, share a sequence homology of greater than 72%;all of the proteins contain a pfam00332 conserved domain and a β-1,3-glucanase active center sequence (LIVM)-X-(LIVMFYW)3-(STAG)-E-(ST)-G-W-P-(ST)-X-G. Furthermore, their phylogenetic relationships are consistent with their traditional classifications. These results were meaningful to reveal the molecular mechanism of R. rugosa pollination incompatibility and improve the theory and techniques of breeding ornamental R. rugosa.
文摘HIV-1 infection requires the expression of CD4＋ molecules in colligation with C-C chemokine receptor type 5 （CCR5） and C-X-C chemokine receptor type 4 （CXCR4） as the major coreceptors. The role of SNP in 3＇ untranslated region ofSDF-1 （SDF1-3 ＇A） and low copy number （CN） of the CCL3L1 gene is reported to confer increased resistance to HIV-1 infection. The aim of the present study was to analyze the combinatorial effect of both the variations in protection towards HIV-1 infection in Indian population. The combinatorial effect of genetic variation in terms of SNP in SDF-1 gene and CCL3L1 CN was investigated in 105 healthy individuals and 78 HIV-I patients. Genotyping of SDF-1 was performed by RFLP-PCR and CCL3L1 by real-time PCR using TaqMan chemistry. The genotype frequency distribution of SDF-1 was found to be （SDF-1/SDF-I： 65.4%, SDF-1/SDF1-3＇A： 29.5% and SDFI-3＇A/SDF1-3＇A- 5.1%） in HIV patients as compared to （SDF-1/SDF-I： 64.8%, SDF-1/SDF1-3＇A： 30.5% and SDF1-3 ＇A/SDF1-3 ＇A： 4.7%） in healthy individuals, whereas a range of 1 to 6 copies per diploid genome was observed for CCL3L1 gene.
