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- Title
结核分枝杆菌Rv3586 结构域基因真核表达载体的构建及鉴定.
- Authors
吴骏杰; 王辛; 宋世杰; 姚庆港; 单炳乙; 王莹; 路延之; 柏银兰
- Abstract
Objective: To construct eukaryotic expression plasmids for three domains of Rv3586. a dinucleotide cyclase of cyclic diadenosine monophosphate (c-di-AMP) of Mycobacterium tuberculosis, and express in COS-7 cells. Methods: Three genes of Rv35S6 domains were amplified by PCR and cloned into eukaryotic expression vector pEGFP-N3, which were identified by colony PCR, plasmid enzyme digestion and DNA sequencing. The recombinant plasmids were transfected into COS-7 cells by liposome. The expression of target genes in COS-7 cells was detected by indirect immunofluorescence. Results: Three genes of Rv3586 domains were amplified by PCR and cloned into eukaryotic expression plasmids respectively, which were confirmed by colony PCR. enzyme digestion and DNA sequencing. And the results of indirect immunofluorescence implied that target genes were expressed in COS-7 cells. Conclusion: The eukaryotic expression plasmids of Rv3586 domains were constructed and expressed in COS-7 cells successfully, which will be helpful for the further research on function and application of M. tuberculosis Rv3 586.
- Publication
Progress in Modern Biomedicine, 2017, Vol 17, Issue 18, p3429
- ISSN
1673-6273
- Publication type
Article
- DOI
10.13241/j.cnki.pmb.2017.18.006