期刊文献+
共找到4,099篇文章
< 1 2 205 >
每页显示 20 50 100
Construction and Expression of Tp0453 Recombinant Protein of Treponema pallidum and Development of Indirect ELISA for Diagnosinq Syphilis 被引量:1
1
作者 刘双全 吴移谋 +1 位作者 赵飞骏 曾铁兵 《Chinese Journal of Sexually Transmitted Infections》 2005年第1期30-34,共5页
Objectives: To clone and express Tp0453 outer membrane protein of Treponema pallidum, and to evaluate its significance in the serodiagnosis of syphilis. Methods: The immuno-dominant epitope of Tp0453 gene was amplif... Objectives: To clone and express Tp0453 outer membrane protein of Treponema pallidum, and to evaluate its significance in the serodiagnosis of syphilis. Methods: The immuno-dominant epitope of Tp0453 gene was amplified from the complete genome of T.pallidum by polymerase chain reactions (PCR), subcloned into the expression vector Pqe32 to generate recombinant plasmid Pqe32/Tp0453, and was then expressed in E. coli M15. The fusion protein was purified with Ni-NTA affinity purification. Indirect ELISA was developed to detect human serum IgG antibody to T. pallidum. Results: The recombinant Tp0453 protein was successfully expressed and purified. The recombinant protein had a molecular weight of approximately 32KDa.Indirect ELISA to the recombinant protein was developed.Sixty control sera were tested by ELISA and yielded a sensitivity of 100% (30/30) and a specificity of 100% (30/30). While testing for T. pallidum in human sera, the sensitivity and specificity of ELISA were 96.8% and 100%, respectively, when compared with TPPA test results. The concordance of results between the ELISA test and the TPPA test was 98.2%. Conclusion: The recombinant Tp0453 outer membrane protein elicited a strong immunoreaction to anti-T.pallidum IgG antibody and has great potential use in ELISA for the serodiagnosis of syphilis. 展开更多
关键词 Treponema pallidum recombinant protein SERODIAGNOSIS enzyme link immunosorbent assay
下载PDF
Induced Expression and Optimal Expression Conditions of Recombinant Alpha-bungarotoxin Gene Fusion Protein
2
作者 贾艳 陈一飞 +3 位作者 胡延春 谢光洪 张乃生 柳增善 《Agricultural Science & Technology》 CAS 2013年第5期716-721,共6页
[Objective] This study aimed to obtain recombinant alpha-bungarotoxin (a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E... [Objective] This study aimed to obtain recombinant alpha-bungarotoxin (a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E coil BL21 (DE3) and BL21 (DE3) plysS host bacteria to identify the optimal engineering strain. Fusion expression of the optimal engineering strain was induced, in order to optimize the induced expression conditions of the soluble fusion protein. [Result] JP-a-BGT was identified as the optimal engineering strain, which could express fusion protein after induced by IPTG. The optimal induced expression conditions of the soluble fusion protein were investigatect JP-a-BGT was incubated at 37 ℃ for 2.5 h and induced with 0.50 mmol4. IPTG for 4 h at 22 ℃, and the expression level of the soluble fusion protein reached 18.42%. [Conclusion] This study laid a solid foundation for the subsequent purification of fusion proteins and the separation and purification of a-BGT. 展开更多
关键词 recombinant alpha-bungarotoxin Fusion protein Expression condition
下载PDF
Immunization of Male Mice with a New Recombinant GnRH Fusion Protein Reduces the Testicular Function 被引量:8
3
作者 FANG Fu-gui YANG Ya-ping +5 位作者 LIU Ya ZHANG Yun-hai TAO Yong WANG Suo-lu PU Yong ZHANG Xiao-rong 《Agricultural Sciences in China》 CAS CSCD 2009年第3期380-385,共6页
The objective of the present study was to evaluate the effect of an immunogenic maltose-binding protein-gonadotropin releasing hormone (GnRH-6-MBP) using genetic engineering. The synthetic mammalian tandem repeated ... The objective of the present study was to evaluate the effect of an immunogenic maltose-binding protein-gonadotropin releasing hormone (GnRH-6-MBP) using genetic engineering. The synthetic mammalian tandem repeated GnRH hexamer gene was inserted into the expression plasmid pMAL-c2x. Recombinant GnRH-6-MBP protein was over- expressed in E.coli strain BL21. Amylose resin with affinity chromatograph was used to purify target protein. The reactiongenicity of fusion protein was identified by indirect enzyme linked immunosorbent assay (ELISA), and the antigenicity and biological effects of GnRH-6-MBP were tested in mice. In the experiment, 20 male Kunming white mice of 20 d old were randomly divided into treatment and control group. Ten mice were immunized with 100 μg GnRH-6-MBP administered subcutaneously (s.c.) thrice at 2-week intervals with GnRH-6-MBP. Mice were sacrificed after 3 weeks following the booster injection, the testis was removed, weighed and measured, and the histological structure was observed. The reactiongenicity of fusion protein to GnRH antibody was much higher than the control. Active immunization against GnRH-6-MBP reduced remarkably (P 〈 0.01) the length and weight of the testis, and shortened the girth and width of the testis (P 〈 0.05), and suppressed testicular spermatogenesis compared to the control mice. These results indicate that the recombinant GnRH-6-MBP acted as a strong immunogen and caused atrophy of the testis. 展开更多
关键词 GNRH recombinant vaccine maltose-binding protein (MBP) IMMUNOCASTRATION
下载PDF
Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line 被引量:7
4
作者 MirjanaMartic EricK.Moses +5 位作者 TimE.Adams DeYiLiu DebraA.Gook ClaireGarrett MarjorieE.Dunlop GordonH.W.Baker 《Asian Journal of Andrology》 SCIE CAS CSCD 2004年第1期3-13,共11页
Aim: To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests. Methods: The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3... Aim: To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests. Methods: The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3 proteins individually and together by co-expression. Presence of these proteins in the culture medium and cell lysate was assessed by Western blotting analysis. The effect of the recombinant proteins on the human AR was assessed. Results: RhZP2 and rhZP3 were secreted into the culture medium, whereas rhZPl was found only in the cell lysate. Interestingly, when all zona pellucida proteins were co-expressed in the same cells, rhZPl was also secreted into the culture medium. However, despite the presence of all three ZP proteins in sufficient concentration and evidence of heavy glycosylation on gel electrophoresis, biological activity to induce the AR was not observed. Conclusion: RhZP1, rhZP2 and rhZP3 were successfully expressed in the human embryonic kidney cell line 293T. It appears that an interaction amongst these proteins may be required for release of rhZPl from the cell. Although this approach is not satisfactory for producing active human ZP proteins, it makes a significant contribution to the understanding of the structural and functional characteristics of the ZP proteins. 展开更多
关键词 acrosome reaction GLYCOSYLATION human cell line recombinant proteins zona pellucida
下载PDF
Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein 被引量:6
5
作者 Yong FU Shen-qing WANG +3 位作者 Ying-peng LIU Guo-peng WANG Jian-ting WANG Shu-sheng GONG 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2008年第4期299-305,共7页
Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of ... Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. Methods: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. Results: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. Conclusion: We have success- fully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies. 展开更多
关键词 recombinant adenovirus vector Viral infection Fetal neural stem cells Green fluorescent protein
下载PDF
Heterotopic ossification after the use of recombinant human bone morphogenetic protein-7 被引量:3
6
作者 Marianthi Papanagiotou Zoe H Dailiana +5 位作者 Theophilos Karachalios Sokratis Varitimidis Michael Hantes Georgios Dimakopoulos Marianna Vlychou Konstantinos N Malizos 《World Journal of Orthopedics》 2017年第1期36-41,共6页
AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7(rhB MP-7) for the treatment of nonunions.METHODS Bone morphogenetic proteins(BMPs) promote bone ... AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7(rhB MP-7) for the treatment of nonunions.METHODS Bone morphogenetic proteins(BMPs) promote bone formation by auto-induction. Recombinant human BMP-7 in combination with bone grafts was used in 84 patients for the treatment of long bone nonunions. All patients were evaluated radiographicaly for the development of heterotopic ossification during the standard assessment for the nonunion healing. In all patients(80.9%) with radiographic signs of heterotopic ossification, a CT scan was performed. Nonunion site palpation and ROM evaluation of the adjacent jointswere also carried out. Factors related to the patient(age, gender), the nonunion(location, size, chronicity, number of previous procedures, infection, surrounding tissues condition) and the surgical procedure(graft and fixation type, amount of rhB MP-7) were correlated with the development of heterotopic ossification and statistical analysis with Pearsons χ~2 test was performed.RESULTS Eighty point nine percent of the nonunions treated with rh BMP-7, healed with no need for further procedures. Heterotopic bone formation occurred in 15 of 84 patients(17.8%) and it was apparent in the routine radiologi-cal evaluation of the nonunion site, in a mean time of 5.5 mo after the rh BMP-7 application(range 3-12). The heterotopic ossification was located at the femur in 8 cases, at the tibia in 6, and at the humerus in οne patient. In 4 patients a palpable mass was present and only in one patient, with a para-articular knee nonunion treated with rhB MP-7, the size of heterotopic ossification affected the knee range of motion. All the patients with heterotopic ossification were male. Statistical analysis proved that patient's gender was the only important factor for the development of heterotopic ossification(P = 0.007). CONCLUSION Heterotopic ossification after the use of rh BMP-7 in nonunions was common but it did not compromise the final clinical outcome in most cases, and affected only male patients. 展开更多
关键词 NONUNION BONE morphogenetic protein recombinant human BONE morphogenetic protein-7 HETEROTOPIC OSSIFICATION Long BONE BONE GRAFT OSTEOINDUCTION
下载PDF
Rat recombinant β-defensin 22 is a heparin-binding protein with antimicrobial activity 被引量:1
7
作者 Hua Diao He-Guo Yu +2 位作者 Fei Sun Yong-Lian Zhang Nongnuj Tanphaichitr 《Asian Journal of Andrology》 SCIE CAS CSCD 2011年第2期305-311,共7页
Approximately 40-50 β-defensins are predominantly expressed in the male reproductive system of mammals. This selective expression raises the question as to the roles of these molecules in innate immunity and fertilit... Approximately 40-50 β-defensins are predominantly expressed in the male reproductive system of mammals. This selective expression raises the question as to the roles of these molecules in innate immunity and fertility in the male reproductive tract. Rat β-defensin 22 is an epididymis-specific β-defensin expressed in segments 12-14 of the epididymis. This protein contains both β-defensin and lectin signature sequences, yet its antimicrobial activity and carbohydrate-binding ability have not been shown. We herein demonstrated the antimicrobial activity of recombinant rat β-defensin 22 against Escherichia coliand Candida albicans. Its lectinJike activity was also investigated by demonstrating its binding ability with heparin beads. This heparin-binding activity implies some potential roles for this defensin in determining the fertilisation capabilities of sperm. 展开更多
关键词 EPIDIDYMIS RAT recombinant protein SPERM
下载PDF
Expression of fluorescent tagged recombinant erythroferrone protein 被引量:1
8
作者 Min Min Than Jetsada Ruangsuriya +1 位作者 Chairat Uthaipibull Somdet Srichairatanakool 《Asian Pacific Journal of Tropical Biomedicine》 SCIE CAS 2018年第7期360-364,共5页
Objective: To produce fluorescent tagged recombinant erythroferrone protein(ERFE_eGFP) for laboratory investigations. Methods: Erythroferrone(ERFE) gene was fused to green fluorescent protein(eGFP) gene and cloned in ... Objective: To produce fluorescent tagged recombinant erythroferrone protein(ERFE_eGFP) for laboratory investigations. Methods: Erythroferrone(ERFE) gene was fused to green fluorescent protein(eGFP) gene and cloned in a pSecTag2Hygro plasmid. The constructed plasmid was amplified in Escherichia coli DH5α and the eGFP-fused ERFE(ERFE_eGFP) protein was expressed in human embryonic kidney(HEK293T) cell line. Results: The plasmid constructed from colony C6 contained ERFE_eGFP with the correct restriction sizes of 4.2 kb and expressed secretory ERFE_eGFP fusion protein(approximately size of 75 kDa) in HEK293T cell line. Conclusions: ERFE_eGFP recombinant protein is successfully expressed as a secretory functional protein and could be sensitively detected using fluorometry. This fusion protein might benefit future applications for localization of cellular ERFE receptors and competitive immunoassay of ERFE concentration. 展开更多
关键词 IRON HEPCIDIN Erythroferrone recombinant protein HEK293T cell
下载PDF
Induction of Chondrogenesis of Adipose-derived Stem Cells by Novel Recombinant TGF-β3 Fusion Protein 被引量:1
9
作者 郑东 但洋 +7 位作者 黄朋 夏天 杨述华 许伟华 杨操 刘国辉 刘先哲 冯勇 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2013年第4期536-542,共7页
Summary: A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 g... Summary: A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells (ADSCs). The recombinant pIRES- EGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFE LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, and the recombinant plasmid of pIRES-EGFP- LAP-MMP-mTGF-β3 was constructed, which was transferred to ADSCs. The ADSCs were cultured and divided in three groups: experimental group (MMP group), negative control group (no MMP) and non-transfection group. The morphological changes were observed microscopically, and the expression of proteoglycan and type II collagen (Col II) was detected by using Alcian blue staining and immuno- histochemistry staining at 7th, 14th and 21st day after culture. The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and se- quencing analysis. The mTGF-β3 fusion protein was successfully expressed after transfection, and in the presence of the MMP, active protein mTGF-β3 was generated, which significantly promoted differ- entiation of ADSCs into chondrocytes and the expression of cartilage matrix. The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes, which would open up prospects for target therapy of cartilage damage repair in future. 展开更多
关键词 adipose-derived stem cells recombinant protein gene clone TGF-Β3 CHONDROGENESIS car-tilage damage target therapy
下载PDF
Immunogenicity and protective efficacy of recombinant M2e.Hsp70c(Hsp70_(359–610)) fusion protein against influenza virus infection in mice 被引量:2
10
作者 Hamidreza Attaran Hassan Nili Majid Tebianian 《Virologica Sinica》 SCIE CAS CSCD 2014年第4期218-227,共10页
New strategies in vaccine development are urgently needed to combat emerging influenza viruses and to reduce the risk of pandemic disease surfacing. Being conserved, the M2 e protein, is a potential candidate for univ... New strategies in vaccine development are urgently needed to combat emerging influenza viruses and to reduce the risk of pandemic disease surfacing. Being conserved, the M2 e protein, is a potential candidate for universal vaccine development against influenza A viruses. Mycobacterium tuberculosis Hsp70(mHsp70) is known to cultivate the function of immunogenic antigen-presenting cells, stimulate a strong cytotoxic T lymphocyte(CTL) response, and stop the induction of tolerance. Thus, in this study, a recombinant protein from the extracellular domain of influenza A virus matrix protein 2(M2e), was fused to the C-terminus of Mycobacterium tuberculosis Hsp70(Hsp70c), to generate a vaccine candidate. Humoral immune responses, IFN-γ-producing lymphocyte, and strong CTL activity were all induced to confirm the immunogenicity of M2 e.Hsp70c(Hsp70359–610). And challenge tests showed protection against H1N1 and H9N2 strains in vaccinated groups. Finally these results demonstrates M2 e.Hsp70c fusion protein can be a candidate for a universal influenza A vaccine. 展开更多
关键词 influenza A virus M2e.Hsp70 recombinant fusion protein universal influenza vaccine
下载PDF
Green Fluorescent Protein Recombinant Nisin as a Probe for Detection of Gram-Positive Bacteria 被引量:1
11
作者 Xiqian Tan Ye Han +1 位作者 Huazhi Xiao Zhijiang Zhou 《Transactions of Tianjin University》 EI CAS 2017年第4期334-339,共6页
A great amount of foodborne pathogens were Gram-positive (G+) bacteria, a threat to public health. In this study, considering the binding ability of nisin towards G+ bacteria and the stable fluorescent ability of... A great amount of foodborne pathogens were Gram-positive (G+) bacteria, a threat to public health. In this study, considering the binding ability of nisin towards G+ bacteria and the stable fluorescent ability of EGFP protein, a fluorescent nisin–EGFP protein probe was constructed by a gene engineering method. Nisin and EGFP were used as the receptor and fluorophore, respectively, to detect G+ bacteria. The nisin and egfp gene were amplified separately according to the sequence published in GenBank using unique primers. The two genes were cloned into a pET-28b(+) vector resulting in a pET-28b(+)–nisin–egfp vector. The vector was transferred into Escherichia coli (E. coli) BL21 (DE3) for expression. The expressed protein was extracted, purified by a Ni–NTA column, and then tested by the SDS-PAGE method to confirm its molecular weight. Listeria monocytogenes (L. monocytogenes), Staphylococcus aureus (S. aureus), and Micrococcus luteus (M. luteus) were used as the representations of G+ bacteria. E. coli O157, representing the gram-negative (G−) bacteria, was used as a negative control. The binding specificity of the recombinant protein was performed on two types of bacteria and then detected through fluorescent microscopy. The results indicated that the nisin–EGFP probe could detect G+ bacteria at 108CFU/mL. © 2017, Tianjin University and Springer-Verlag Berlin Heidelberg. 展开更多
关键词 ANTIBIOTICS Bins Cloning Escherichia coli Fluorescence Genes Health risks PATHOGENS Probes proteins recombinant proteins
下载PDF
Production of spike and nucleocapsid recombinant proteins of porcine epidemic diarrhea virus for antibody detection by ELISA 被引量:1
12
作者 Anchalee Srijangwad Dachrit Nilubol +3 位作者 Wanchai Chongcharoen Waranyoo Phoolcharoen Taksina Chuanasa Angkana Tantituvanon 《Asian Journal of Pharmaceutical Sciences》 SCIE CAS 2016年第1期85-86,共2页
Porcine epidemic diarrhea (PED), a devastating enteric disease in pigs, is caused by PEDvirus (PEDV)(1)Reduced severity of clinical diseases was reported to associate with neutralizing antibody titers in colostrum. Ho... Porcine epidemic diarrhea (PED), a devastating enteric disease in pigs, is caused by PEDvirus (PEDV)(1)Reduced severity of clinical diseases was reported to associate with neutralizing antibody titers in colostrum. However, viral neutralization assay(VN) is laborious and not suitable for routine diagnosis. Spike protein plays an important role in stimulating neutralizing antibody that might be suitable for PEDV diagnosis. 展开更多
关键词 recombinant protein SPIKE NUCLEOCAPSID PORCINE EPIDEMIC DIARRHEA ELISA
下载PDF
CONSTRUCTION, EXPRESSION AND BIOLOGICAL ASSESSMENT OF BPI_(23)-Fcγ1 RECOMBINANT PROTEIN PROKARYOTIC EXPRESSION VECTOR 被引量:7
13
作者 安云庆 管远志 +1 位作者 柯岩 杨贵贞 《Chinese Medical Sciences Journal》 CAS CSCD 2002年第3期140-147,共8页
关键词 pBV BPI600 Fcγ1700 recombinant expression vector BPI23 Fcγ1 recombinant protein Objective. To construct pBV BPI600 Fcγ1700 recombinant expression vector to transform it into Escherichia coli DH5α and to induce the expression of BPI2
下载PDF
Susceptibility to AcMNPV and Expression of Recombinant Proteins by a Novel Cell Clone Derived from a Trichoplusia ni QAU-BTI-Tn9-4s Cell Line 被引量:1
14
作者 Ming Shan Shi-ying Zhang +2 位作者 Lei Jiang Ming Ma Guo-xun Li 《Virologica Sinica》 SCIE CAS CSCD 2011年第5期297-305,共9页
It is well known that Tn5B1-4 (commercially known as the High Five) cell line is highly susceptible to baculovirus and provides superior production of recombinant proteins when compared to other insect cell lines.But ... It is well known that Tn5B1-4 (commercially known as the High Five) cell line is highly susceptible to baculovirus and provides superior production of recombinant proteins when compared to other insect cell lines.But the characteristics of the cell line do not always remain stable and may change upon continuous passage.Recently an alphanodavirus,named Tn5 Cell Line Virus (or TNCL Virus),was identified in High Five cells in particular.Therefore,we established a new cell line,QB-Tn9-4s,from Trichoplusia ni,which was determined to be free of TNCL virus by RT-PCR analysis.In this paper,we describe the development of a novel cell clone,QB-CL-B,from a low passage QB-Tn9-4s cell line and report its susceptibility to AcMNPV,and the level of recombinant protein production.This cell clone was similar to its parental cells QB-Tn9-4s and Tn5B1-4 cells in morphology and growth rate;although it also showed approximately the same responses to AcMNPV infection and production of occlusion bodies,there were higher levels of recombinant protein production in comparison to QB-Tn9-4s (parental cells) and High5 cells. 展开更多
关键词 Cell line Insect virus recombinant protein expression RT-PCR TNCL virus
下载PDF
Recombinant human bone morphogenetic protein-7 expressed from CHO cells possessing the activity of bone-induced in vitro 被引量:1
15
作者 LI Xiaoyan WANG Hao +4 位作者 YANG Yang TAN Min XUE Jingya NI Haidong GUO Yajun 《脊柱外科杂志》 2006年第3期159-162,182,共5页
Objective To express the recombinant human bone morphogenetic protein-7 (rhBMP-7) in Chinese hamster ovary (CHO) cells and to establish the in vitro biological activity assay of rhBMP-7. Methods Human BMP-7 cDNA was s... Objective To express the recombinant human bone morphogenetic protein-7 (rhBMP-7) in Chinese hamster ovary (CHO) cells and to establish the in vitro biological activity assay of rhBMP-7. Methods Human BMP-7 cDNA was subcloned into pcDNA3.1 mammalian expression vector and transfected to CHO cells by using the lipofectin transfection method. BMP-7 expression cell culture supernatants were harvested and purified for target protein. To analyze the bioactivity of the secreted rhBMP-7, a novel in vitro assay was established by measuring its alkaline phosphatase (ALP) stimulating of osteoblast cell line, W-20-17. Results BMP-7 stably expressing cell clone was selected, which secreted mature disulfide-linked homodimer form of hBMP-7 and had an apparent molecular weight of 36kDa. rhBMP-7 with >95% purity was obtained using 3 step chromatography method. Bioactivity assay showed that the purified protein specifically stimulated W-20-17 cell producing ALP, with a 4-fold increase of ALP activity at 100ng/ml or more, and the EC50 of 15.6ng/ml. Conclusion Purified rhBMP-7 from this CHO expression system has significant biological activity in induction of osteoblast phenotype, which demonstrates potential bone regeneration activity. 展开更多
关键词 bone morphogenetic proteins recombinant proteins alkaline phosphatase CHO cells in vitro gel chromatography
下载PDF
Expression and purification of the recombinant outermembrane protein Tp0453 of Treponema pallidum and its characterization of immuno-competence 被引量:3
16
作者 SHUANGQUANLIU YIMOUWU FEIJUNZHAO TIEBINGZENG WEIGUOYIN 《Journal of Microbiology and Immunology》 2005年第1期47-52,共6页
To clone and express the recombinant outer membrane protein Tp0453 of Treponema pallidum and to analyze the immuno-reactivity and immunogenicity of the expressed protein, the immuno-dominant epitope of the Tp0453 was ... To clone and express the recombinant outer membrane protein Tp0453 of Treponema pallidum and to analyze the immuno-reactivity and immunogenicity of the expressed protein, the immuno-dominant epitope of the Tp0453 was amplified from the complete genome of T.pallidum by PCR, subcloned into expression vector pQE32 to generate the recombinant plasmid pQE32/Tp0453, then expressed in E.coli M15 and analyzed by SDS/PAGE and Western blotting. The fusion protein expressed was purified with Ni-NTA affinity chromatography. Its immuno-reactivity was assayed by indirect ELISA, and the immunogenicity was determined by immunization with this fusion protein in New Zealand rabbits. In the present study, a fusion protein of molecular weight about 32 kDa was obtained. As demonstrated by Western blotting, the recombinant protein could react specifically with positive IgG sera of patients with syphilis, and the antibodies against T.pallidum in human sera were successfully detected by indirect ELISA. Both the sensitivity and specificity of ELISA based on the Tp0453 fusion protein as were 100% (30/30) when detected with control sera. In comparison with the results of IgG ELISA with those of TPPA. It was found that the sensitivity of ELISA was 96.8% and the specificity was 100%. The difference of ELISA and TPPA was not significant, and the concordance of results between ELISA and TPPA was 98.2%. In addition, specific humoral responses could be elicited by immunization with the recombinant fusion protein in New Zealand rabbits with a specific antibody titer of 1∶1280 after 3 successive doses of immunization. These results demonstrate that the expressed recombinant fusion protein shows excellent immuno-competence and provide foundation to develop a quick diagnostic kid applied to detect the presence of T.pallidum infections. 展开更多
关键词 Treponema pallidum recombinant protein TP0453 Immuno-competence
下载PDF
Use of recombinant human bone morphogenetic protein-2 in spine surgery 被引量:5
17
作者 Marios Lykissas Ioannis Gkiatas 《World Journal of Orthopedics》 2017年第7期531-535,共5页
Bone morphogenetic proteins are osteoinductive factors which have gained popularity in orthopaedicsurgery and especially in spine surgery. The use of recombinant human bone morphogenetic protein-2 has been officially ... Bone morphogenetic proteins are osteoinductive factors which have gained popularity in orthopaedicsurgery and especially in spine surgery. The use of recombinant human bone morphogenetic protein-2 has been officially approved by the United States Food and Drug Administration only for single level anterior lumbar interbody fusion, nevertheless it is widely used by many surgeons with off-label indications. Despite advantages in bone formation, its use still remains a controversial issue and several complications have been described by authors who oppose their wide use. 展开更多
关键词 recombinant human BONE morphogenetic protein-2 SPINE FUSION BONE GRAFT Yale UNIVERSITY Open Data project
下载PDF
Development of an Indirect ELISA for Detection of T.solium Based on Recombinant 18 kDa Protein
18
作者 WU Guo-hua ZHENG Ya-dong +5 位作者 JIA Wan-zhong ZHANG Shao-hua JING Zhi-zhong LUO Xue-nong LIU Shi-quan CAI Xue-peng 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期87-92,共6页
The gene encoding the 18 kDa protein of Taenia solium metacestodes was amplified by RT-PCR and cloned into the pGEM-T vector for sequencing.The recombinant plasmid named pGEX-CE18 was constructed and transformed into ... The gene encoding the 18 kDa protein of Taenia solium metacestodes was amplified by RT-PCR and cloned into the pGEM-T vector for sequencing.The recombinant plasmid named pGEX-CE18 was constructed and transformed into E.coli BL21 for in vitro expression.SDS-PAGE and Western blot were employed for analyzing the recombinant protein,which was then used for development of an indirect ELISA for detection of anti-cysticercosis antibodies.The results showed that the recombinant protein of interest was 35 kDa in size,accounting for 28%of total bacteria proteins,and reacted with positive sera against cysticercosis.Using the newly-constructed indirect ELISA and a commercially available ELISA kit,paired analyses of 178 serum samples indicated that the concordant rate was 98.83%and the ELISA exhibited good specificity and sensitivity,supporting its utility and application for diagnosis of cysticercosis. 展开更多
关键词 CYSTICERCUS cellulosae 18 kDa protein E.COLI EXPRESSION recombinant protein ELISA
下载PDF
Evaluation of Six Recombinant Proteins for Serological Diagnosis of Lyme Borreliosis in China
19
作者 LIU Wei LIU Hui Xin +3 位作者 ZHANG Lin HOU Xue Xia WAN Kang Lin HAO Qin 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2016年第5期323-330,共8页
Objective In this study, we evaluated the diagnostic efficiency of six recombinant proteins for the serodiagnosis of Lyme borreliosis (LB) and screened out the appropriate antigens to support the production of a Chi... Objective In this study, we evaluated the diagnostic efficiency of six recombinant proteins for the serodiagnosis of Lyme borreliosis (LB) and screened out the appropriate antigens to support the production of a Chinese clinical ELISA (enzyme-linked immunosorbent assay) kit for LB. Methods Six recombinant antigens, Fla B.g, OspC B.a, OspC B.g, P39 B.g, P83 B.g, and VlsE B.a, were used for ELISA to detect serum antibodies in LB, syphilis, and healthy controls. The ELISA results were used to generate receiver operating characteristic (ROC) curves, and the sensitivity and specificity of each protein was evaluated. All recombinant proteins were evaluated and screened by using logistic regression models. Results Two IgG (VISE and OspC B.g) and two IgM (OspC B.g and OspC B.a) antigens were left by the logistic regression model screened. VIsE had the highest specificity for syphilis samples in the IgG test (87.7%, P〈0.05). OspC B.g had the highest diagnostic value in the IgM test (AUC=0.871). Interactive effects between OspC B.a and Fla B.g could reduce the specificity of the ELISA. Conclusion Three recombinant antigens, OspC B.g, OspC B.a, and VisE B.a, were useful for ELISAs of LB. Additionally, the interaction between OspC B.a and Fla B.g should be examined in future research. 展开更多
关键词 Lyme borreliosis recombinant proteins Serological diagnosis Logistic regression ROC
下载PDF
The development of a recombinant hybrid protein of Plasmodium falciparum and analysis of its antigenicity and protectivity
20
作者 李全贞 李英杰 +3 位作者 谢毅 任大明 毕惠祥 徐秉锟 《Journal of Medical Colleges of PLA(China)》 CAS 1994年第3期161-169,共9页
A hybrid gene encoding several putative protective epitopes from erythrocytic stage antigens (MSA1,MSA2 and RESA) of P. falciparum as well as exgenous T cell enhancer epitopes from interleukin-1 and tetanus toxin was ... A hybrid gene encoding several putative protective epitopes from erythrocytic stage antigens (MSA1,MSA2 and RESA) of P. falciparum as well as exgenous T cell enhancer epitopes from interleukin-1 and tetanus toxin was synthesized chemically. This gene (named HGFC) was cloned and connected with another hybrid gene (HPFA) synthesized previously to make a bigger hybrid gene (HGFCAC). HGFC and HGFCAC were cloned in an expression vector pWR450-1 and transformed into E. Coli JM109. The engineered bacteria could express fusion proteins with molecular weights of 65 and 77 kDa after inducing with isopropylthio-β-D-galactoside (IPTG). The expression rate was about 35% of total bacterial proteins. The expressed products showed sepcific immunological reaction with rabbit antibodies against P. falciparum peptide Glu-Glu-Asn-Val-Glu-His-Asp-Ala (EENVEHDA)by Western blotting. The fusion proteins were pruified by precipitation with amonium sulfate. gel filtration and ion-exchange chromatography and the purity was 82%. The purified protein reacted specifically with mouse immune serum against falciparum blood stage antigens by dot enzyme-linked immunosorbent assay (dot-ELISA).The fusion protein was emulsified with Freund's complete adjuvant (FCA) and used to immunize rabbits. The immune serum can recognize P. falciparum erythrocytic stage antigens of Fcc-1/HN strain and Yunnan strain and had weak cross reaction with P. vivax,but had no reaction with P. cynomlogi and P. berghei antigens. The protective effect of the antibody was tested by in vitro inhibition test to cultured falciparum parasites. Preliminary results indicated that the immune sera could effectively reduce the invasion rates of the parasites to red blood cells and inhibit the growth of the in vitro cultured falciparum parasites. The inhibitory capacity of the immune sera to parasite invasion is enhanced as the amount of the sera increases and the incubation time of the sera with the parasites is prolonged.It was shown that after 72 h incubation at 20% concentration with the parasites, the serum can suppress the multiplication of P.falciparum growth in vitro to a level of 80%.The immune sera caused dispersion of the parasite cytoplasm,atrophy of parasites,agglutination of free merozoites and degeneration of schizonts. 展开更多
关键词 PLASMODIUM FALCIPARUM recombinant protein HYBRID antigen inhibition test in vitro malarial vaccine
下载PDF
上一页 1 2 205 下一页 到第
使用帮助 返回顶部