检索结果-南通市图书馆

Expression and purification of the recombinant outermembrane protein Tp0453 of Treponema pallidum and its characterization of immuno-competence

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Journal of Microbiology and Immunology 2005年第1期3卷 47-52页

作者： SHUANGQUANLIU YIMOUWU FEIJUNZHAO TIEBINGZENG WEIGUOYIN InstituteofPathogenicBiology MedicalCollegeNanhuaUniversityHengyangP.R.China

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 *** by PCR, subcloned into expression vector pQE32 to generate the recombinant plasmid pQE32/Tp0453, then expressed in *** 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 *** 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 *** infections.

关键词： Treponema pallidum recombinant protein Tp0453 Immuno-competence

Rat recombinant β-defensin 22 is a heparin-binding protein with antimicrobial activity

在线全文

学校读者我要写书评

暂无评论

Asian Journal of Andrology 2011年第2期13卷 305-311页

作者： Hua Diao He-Guo Yu Fei Sun Yong-Lian Zhang Nongnuj Tanphaichitr NPFPC Key Laboratory of Contraceptives and Devices Shanghai Institute of Planned Parenthood Research Shanghai 200032 China Laboratory for Reproductive Biology School of Life Science University of Science and Technology of China Hefei 230027 China Shanghai Key La boratory for Molecular Andrology State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Chronic Disease Program Ottawa Hospital Research Institute Ottawa Ontario KIY 4E9 Canada

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.

关键词： epididymis rat recombinant protein sperm

Induction of Chondrogenesis of Adipose-derived Stem Cells by Novel recombinant TGF-β3 Fusion protein

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Journal of Huazhong University of Science and Technology(Medical Sciences) 2013年第4期33卷 536-542页

作者：郑东但洋黄朋夏天杨述华许伟华杨操刘国辉刘先哲冯勇 Department of Orthopedics Union HospitalTongji Medical CollegeHuazhong University of Science and Technology

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.

关键词： adipose-derived stem cells recombinant protein gene clone TGF-β3 chondrogenesis car-tilage damage target therapy

Development of a recombinant pB602L-based indirect ELISA assay for detecting antibodies against African swine fever virus in pigs

在线全文

学校读者我要写书评

暂无评论

Journal of Integrative Agriculture 2022年第3期21卷 819-825页

作者： WANG Peng-fei WANG Ming SHI Zhi-bin SUN Zhen-zhao WEI Li-li LIU Zai-si WANG Shi-da HE Xi-jun WANG Jing-fei State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069P.R.China

African swine fever(ASF),caused by the African swine fever virus(ASFV),is a devastating disease of domestic and wild *** is no effective vaccine,and the control of the disease relies mainly on surveillance and early detection of infected ***,serological assays,such as ELISA,have been developed mainly based on recombinant structural viral proteins of ASFV,including p72,p54,and ***,the antibodies against these proteins do not provide efficient protection against ASFV infection in ***,new serological assays that can be applied for clinical diagnosis and evaluating serological immune response in vaccinated pigs are still *** this study,we expressed and purified a recombinant p B602 L *** purified p B602 L protein was then used as an antigen to develop an indirect ELISA *** assay has no cross-reaction with the anti-sera against the 15 most common pig pathogens in China,such as classical swine fever virus,pseudorabies virus,and porcine *** assay and a commercial ELISA kit were then used to detect 60 field pig serum samples,including an unknown number of antiASFV *** coincidence of the two assays was 95%.Furthermore,the p B602 L-based ELISA was employed to test the antibody responses to the seven-gene-deleted ASFV strain HLJ/18-7 GD in *** results showed that the antibody levels in all vaccinated pigs,starting from the 10 th day post-inoculation,have increased continuously during the observation period of 45 *** results indicate that this p B602 L-based indirect ELISA assay can be employed potentially in the field of ASFV diagnosis.

关键词： African swine fever virus pB602L recombinant protein indirect ELISA assay pig

Effect of artmether,hemin and Fe^(3+) on recombinant lactate dehydrogenase from Schistosoma japonicum

在线全文

学校读者我要写书评

暂无评论

Asian Pacific Journal of Tropical Medicine 2010年第12期3卷 930-933页

作者： Gang Lu Xuchu Hu Can Huang Yajun Lu Lixian Wu Lihua Li Jing Xu Xinbing Yu Department of Pathogen Biology Hainan Medical University Department of Parasitology Zhongshan School of MedicineSun Yat-Sen University

Objective:To explore antischitosome effects of artemether,hemin and Fe on S/***: Enzyme activity of rS/LDH was assayed in the standard reaction system by adding different concentration of reagents(0.00-0.10 mM artemether,0.00-0.02 mM hemin,0.00-0.50 mM Fe^(3+)). Same solvents of the each reagent were used as ***:There was no enzyme activity inhibition observed at 0.10 mM artemther:obivious inhibition for lactate oxidation reaction and pyruvate reduction reaction were detected at 0.002 mM and 0.004 mM of hemin,respectively: comparing with that of the control(P<0.05).The relative enzymatic activity inhibitions for pyruvate reduction reaction and lactate oxidation reaction at 0.02 mM hemin were 93.48%and 100.00%,respectively,comparing with that of the control(P<0.01):both pyruvate reduction and lactate oxidation reaction were inhibited completely at 0.50 mM Fe^(3+),comparing with that of the control(P<0.01).Conclusions:The results implied that SjLDH was not the direct molecular target of *** and Fe are inhibitors of SjLDH.

关键词： Schistosoma japonicum Lactate dehydrogenase recombinant protein Artemether Hemin Fe^(3+)

Plant-produced recombinant SARS-CoV-2 receptor-binding domain;an economical,scalable biomaterial source for COVID-19 diagnosis

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Biomaterials Translational 2021年第1期2卷 43-49页

作者： Kaewta Rattanapisit Gorawit Yusakul Balamurugan Shanmugaraj Kittinop Kittirotruji Phassorn Suwatsrisakul Eakachai Prompetchara Suthira Taychakhoonavud Waranyoo Phoolcharoen Baiya Phytopharm Co. Ltd.BangkokThailand School of Pharmacy Walailak UniversityNakhon Si ThammaratThailand Centre of Excellence in Vaccine Research and Development(Chula Vaccine Research Centre Chula VRC)Faculty of MedicineChulalongkorn UniversityBangkokThailand Department of Laboratory Medicine Faculty of MedicineChulalongkorn UniversityBangkokThailand Department of Social and Administrative Pharmacy Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkokThailand Research Unit for Plant-produced Pharmaceuticals Chulalongkorn UniversityBangkokThailand Department of Pharmacognosy and Pharmaceutical Botany Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkokThailand

The outbreak of the novel coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),spread rapidly causing a severe global health *** standard COVID-19 diagnosis relies heavily on molecular tests to detect viral RNA in patient samples;however,this method is costly,requires highly-equipped laboratories,multiple reagents,skilled laboratory technicians,and takes 3-6 hours to *** overcome these limitations,we developed a plant-based production platform for the SARS-CoV-2 receptor-binding domain as an economical source of detection reagents for a lateral-flow immunoassay strip(LFIA)which is suitable for detection of IgM/IgG antibodies in human ***,we validated the plant-produced SARS-CoV-2 receptor-binding domain-based LFIA as a useful diagnostic tool for COVID-19.A total of 51 confirmed COVID-19 serum samples were tested using the LFIA,and the obtained results were consistent with those from polymerase chain reaction assays,while providing sensitivity and specificity of 94.1%and 98%,*** developed LFIA is rapid,scalable,user-friendly,and relatively inexpensive with a simple test procedure,making it useful for the routine monitoring of COVID-19 in clinical *** study was approved on March 19,2020 by the Ethics Committee of the Faculty of Medicine,Chulalongkorn University(COA No.354/2020 and IRB No.236/63).

关键词： COVID-19 lateral flow immunoassay Nicotiana benthamiana point-of-care testing recombinant protein SARS-CoV-2

Secretory Expression of recombinant Diphtheria ToxinMutants in B. Subtilis

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Journal of Huazhong University of Science and Technology(Medical Sciences) 1999年第4期19卷 253-256页

作者：周建华 Department of Pediatrics Tongji Hospital Tongji Medical University Wuhan 430030

Three diphtheria toxin (DT) mutants CRM-197, DT-del (148) and DT-El48S-K516A-F530A were cloned in B- Subtilis plasmid PSM604 under the subtilisin signal sequence. The expression was effective in both SMS300 and SMS118, but higher yield of 7. 1 mg/L was observed in SMS300 compared with 2. 1 mg/L in SMS118. Western blot showed that the recombinant protein could be effectively secreted into the culture medium as a 58 ku peptide, and could be de-graded into two peptides of 37ku and 21ku.

关键词： B. Subtilis diphtheria toxin Western blot recombinant protein gene expression mutation

cDNA Cloning, Prokaryotic and Eukaryotic Expression and Characterization of Porcine Leukemia Inhibitory Factor

在线全文

学校读者我要写书评

暂无评论

Chemical Research in Chinese Universities 2006年第2期22卷 145-149页

作者： LI Ming-tang JIANG Yong WANG Shu-mei LI Yong-ming WANG Fang HOU Xia YANG Hong MA Tong-hui Membrane Channel Research Laboratory Northeast Normal University Changchun 130024 P. R. China Department of Anesthesia Second Teaching Hospital Jilin University Changchun 130041 P. R. China

Molecular cloning of the porcine leukemia inhibitor factor（pLIF） has not been reported. A full-length eDNA encoding pLIF was cloned, expressed and characterized. The full-length porcine LIF cDNA encodes a 202 amino acid protein that has an 84% sequence identity to mouse LIF and 86% sequence identity to human LIF. The deduced amino acid sequence of a pLIF protein contains six conserved consensus N-linked glycosylation sites and six cysteine groups to form potential disulfide bonds. The pLIF was expressed in E coli, as a mature form, and in CHO cells as a secreted form. Both the forms of the recombinant pLIFs can maintain murine embryonic stem cells in an undifferentiated state in a culture. The recombinant pLiFs will be useful in establishing a long-term culture of stable pluripotent porcine embryonic stem cells for further manipulation.

关键词： Leukemia inhibitor factor Porcine Cloning recombinant protein Embryonic stem cell Pluripotency

Molecular cloning and characterization of a cotton phosphoenolpyruvate carboxylase gene

在线全文

学校读者我要写书评

暂无评论

Progress in Natural Science:Materials International 2008年第5期18卷 539-545页

作者： Zhixin Qiao, Jin-Yuan Liu Laboratory of Molecular Biology and protein Science Laboratory of the Ministry of Education, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China Laboratory of Molecular Biology and protein Science Laboratory of the Ministry of Education Department of Biological Sciences and Biotechnology Tsinghua University Beijing 100084 China

Phosphoenolpyruvate carboxylase (PEPC) plays diverse physiological functions during plant development. In this study, a new phos- phoenolpyruvate carboxylase gene GhPEPC2 is isolated from cotton (Gossypium hirsutum cv. zhongmian 35) by RACE-PCR. The cloned cDNA of GhPEPC2 is 3364 bp in length, and has an open reading frame of 2913 bp, encoding for 971 putative amino acids with a calculated molecular mass of 110.6 kD and pI of 5.56. The deduced amino acid sequence of GhPEPC2 shares high similarity with other reported plant PEPCs. Southern blot analysis indicates that the cotton PEPC exists as a small gene family and the GhPEPC2 might have two copies in the cotton genome. The semi-quantitative RT-PCR reveals that GhPEPC2 constitutively expresses in all the tissues of cot- ton and accumulated highly in roots, owers and embryos but relatively low in stems and bers. In addition, the recombinant GhPEPC2 has been puri ed by expressing it in Escherichia coli and the catalytic properties of it were also investigated. The results showed that GhPEPC2 is a typical C3 PEPC with a higher Km (83.6 lM) and lower Vmax (8.0 lmol min 1 mg 1) compared with the C3 PEPCs pre- viously reported.

关键词： Phosphoenolpyruvate carboxylase recombinant protein Southern blot Cotton