Cloning and expression of nucleocapsid protein gene of TGEV HB06 strain |
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Authors: | Jinghui Fan Yuzhu Zuo Yuelan Zhao Tanqing Li Xiaobo Zhang |
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Institution: | (1) College of Animal Science and Veterinary Medicine, Agricultural University of Hebei, Baoding, 071001, China;(2) College of Traditional Chinese Veterinary Medicine, Agricultural University of Hebei, Dingzhou, 073000, China |
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Abstract: | The nucleocapsid protein gene of transmissible gastroenteritis virus, 1 149 bp in length, was amplified by RT-PCR from isolated
strain HB06 and cloned into pMD18-T. Sequence comparison with other transmissible gastroenteritis virus (TGEV) strains selected
from the Gene Bank revealed that the homology of N gene complete sequence shares more than 97% in nucleotide. N gene was cloned into BamHI and EcoRI multiple cloning sites of the prokaryotic expression vector pET 20 b, and named pETN. After being induced by isopropyl-β-D-thiogalactopyranoside (IPTG), the recombinant nucleocapsid protein was expressed. The result of SDS-PAGE and Western-blot
showed that the recombinant nucleocapsid protein was 47 kDa and had strong positive reactions with TGEV-specific antibody. |
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Keywords: | transmissible gastroenteritis virus N gene cloning expression |
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