Prokaryotic Expression and Immunoreactivity of Different Encoding Regions of the Spike Gene of Porcine Transmissible Gastroenteritis Virus

doi:10.7668/hbnxb.2014.06.012

Abstract

Abstract: To identify immunoreactivity of different encoding regions of the spike (S) gene of Transmissible gastroenteritis virus (TGEV),three encoding regions containing four antigenic sites C,B,D and A located at the N-terminus of S protein was analyzed and expressed by prokaryotic expression system.The fragments TCB (containing antigenic sites C and B,411 bp),TD (containing antigenic site D,441 bp) and TA (containing antigenic site A,456 bp) were amplified by PCR.After cloning and sequence,the fragments were inserted into the prokaryotic expression vector pET-32a respectively.The fusion proteins yielded by TCB,TD and TA recombinant expression plasmids were 34.6,35.1,36.1 kDa respectively.The expressed proteins showed strong positive reaction with TGEV-infected swine serum and His tag antibody by Western Blot.These results indicated that the three encoding regions of S gene had a strong immunoreactivity with TGEV positive antiserum,which will play an essential role in identifying immunogenicity and immunological protection of different encoding regions of the S protein of TGEV.

Key words: Porcine transmissible gastroenteritis virus, Spike gene, Prokaryotic expression, Immunoreactivity

CLC Number:

Q786
S828

ZHOU Hong-zhuan, TAN Xiang-jie, YANG Bing, XU Fu-zhou, MAN Kun, SU Xia, ZHANG Xiao-dong, LIU Jue, LUO Xu-gang, WANG Jin-luo. Prokaryotic Expression and Immunoreactivity of Different Encoding Regions of the Spike Gene of Porcine Transmissible Gastroenteritis Virus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(6): 63-67. doi: 10.7668/hbnxb.2014.06.012.

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