伽氏疏螺旋体尚志株P66基因的原核表达与鉴定

doi:10.16431/j.cnki.1671-7236.2017.01.030

Abstract

Abstract:

In order to obtain the P66 protein from Borrelia garinii (B. garinii) SZ, the first strand cDNA was synthesized based on the total RNA extracted from B. garinii SZ, and then the targeted P66 gene was amplified by PCR. The fragment was linked into the pET-30a(+) vector, and transformed into Escherichia coli BL21(DE3). After identified by PCR, double restriction enzyme digestion, and nucleotide sequencing, the recombinant protein was expressed and purified. Polyclonal antibody was then prepared from New Zealand rabbit immunized with purified recombinant protein. The recombinant protein was about 70 ku in size confirmed by SDS-PAGE, and Western blotting analysis indicated that the recombinant P66 protein could recognize the mouse monoclonal anti-His-tag, positive sera of spirochete from mouse, and anti-P66 polyclonal antibody. Additionally, the anti-P66 polyclonal antibody could recognize native P66 protein. In this study, we successfully expressed the recombinant P66 protein and obtained the anti-P66 polyclonal antibody, which provided the foundation for further functional studies of P66 protein from B. garinii SZ.

Key words: Borrelia garinii SZ; P66; prokaryotic expression; polyclonal antibody

CLC Number:

S852.63

YU Pei-fa, LIU Zhi-jie, NIU Qing-li, YANG Ji-fei, WANG Zhen-guo, ZHAI Bin-tao, PAN Yu-ping, GUAN Gui-quan, LUO Jian-xun, YIN Hong. Prokaryotic Expression and Identification of P66 Gene from Borrelia garinii SZ[J]. , 2017, 44(1): 214-220.

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Prokaryotic Expression and Identification of P66 Gene from Borrelia garinii SZ

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