Vero E6细胞中B23.1基因的扩增及其原核表达

›› 2012, Vol. 39 ›› Issue (3): 33-36.

• 生物技术 • Previous Articles Next Articles

Amplification and Prokaryotic Expression of B23.1 Protein Gene in Vero E6 Cell

LV Mao-jie^1,2, CHEN Jian-fei¹, SHI Hong-yan¹, FENG Li¹

1. Division of Porcine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;2. Ringpu (Daoding) Biological Pharmaceutical Co.Ltd., Baoding 071000, China

Received:2011-08-01 Revised:1900-01-01 Online:2012-03-20 Published:2012-03-20

Abstract

Abstract: A fragment containing B23.1 protein gene of Vero E6 cell was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using the upper/lower primers each including the recognition sequence for BamH Ⅰ and Xho Ⅰ respectively. The results showed that the ORF of B23.1 gene was composed of 901 bp, deletion and insertion were not found. By digestion with restriction enzymes BamH Ⅰ and Xho Ⅰ, the B23.1 gene were cloned into pGEX-6p-1 vector to construct recombinant plasmid named pGEX-6p-B23.1. The recombinant plasmid was transformed into E.coil BL21(DE3) and induced with IPTG. The results of SDS-PAGE assay showed that the protein was largely expressed,the GST-tagged protein was purified. Western blotting analysis showed that the target protein had a good reactivity with B23 monoclonal antibody (mouse).

Key words: Vero E6 cell; B23.1 gene; recombinant protein; reactivity

CLC Number:

Q786

LV Mao-jie;CHEN Jian-fei;SHI Hong-yan;FENG Li. Amplification and Prokaryotic Expression of B23.1 Protein Gene in Vero E6 Cell[J]. , 2012, 39(3): 33-36.

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Amplification and Prokaryotic Expression of B23.1 Protein Gene in Vero E6 Cell

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