تیمورپور, رقیه ، مشکات, زهرا ، رشیدیان, سمیرا (1396) ایزولاسیون، کلونینگ و آنالیز مولکولی ژن های TB10.4 ,Ag85aاز مایکوباکتریوم توبرکلوزیس. در: 18th international Iranian congress of microbiology, Aug 29-31 , 2017, Tehran - Iran.
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عنوان انگليسی
Isolation, cloning and molecular analysis of ag85a and tb10.4 genes from Mycobacterium tuberculosis
خلاصه انگلیسی
Background: Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis.Materials and Methods: tb10.4 fragment was amplified by PCR and the product was digested with restriction enzymes. Next, it was cloned into the pcDNA3.1+ plasmid. The ag85a gene and pcDNA3.1+/tb10.4 plasmid were digested by EcoRI and BamHI restriction enzymes. Constructed vector was sequenced. The molecular analysis was done using bioinformatics software. New chimeric vector containing ag85a-tb10.4 genes were purified. Expression of pcDNA3.1+/tb10.4-ag85a plasmid was confirmed in eukaryotic cells. Results: Fragments of 297 bp for tb10.4 and 1017 bp for ag85a were observed in agarose gel electrophoresis. Alignment of ag85a-tb10.4 genome sequence with reference genes in GenBank showed exact identities that indicate correction of all cloning procedures. Transfection of eukaryotic cells with pcDNA3.1+/tb10.4-ag85a vector and existence of tb10.4-ag85a fusion gene were both confirmed with RT-PCR.Conclusion: In this study, tb10.4 and ag85a genes were isolated from Mycobacterium tuberculosis H37Rv strain and cloned into pcDNA3.1+. Also, the capability of constructed vector in producing fusion ag85a-tb10.4 protein was confirmed with RT-PCR. pcDNA3.1+/tb10.4-ag85a vector can be used for further studies in future.
نوع سند : | موضوع کنفرانس یا کارگاه (پوستر ) |
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زبان سند : | انگلیسی |
نویسنده اول : | رقیه تیمورپور |
موضوعات : | QW میکروب شناسی و ایمنی شناسی |
بخش های دانشگاهی : | دانشكده پزشكي > گروه علوم پایه > بخش انگل شناسی |
کد شناسایی : | 9807 |
ارائه شده توسط : | دکتر رقیه تیمورپور |
ارائه شده در تاریخ : | 23 بهمن 1396 07:24 |
آخرین تغییر : | 21 فروردین 1397 09:00 |
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