Cloning of 3D of Foot-and-mouth Disease Virus Strain AF72 and Its Expression in Escherichia coli

doi:10.7668/hbnxb.2008.06.007

Abstract

Abstract: The 3D polymerase gene of Foot-and-mouth disease virus strain AF72 was amplified by reverse transcription polymerase chain reaction(RT-PCR),The purified PCR products were cloned into pGEM-T easy Vectors, and the nu- cleotide sequences were highly homologous to other four reference strains from GenBank. The target gene was cloned into the prokaryotic expression vector pET-30a(+), then recombinant expression plasmid pET- 3D was transformed into E. coli BL21(DE3) competent cells, the target protein was highly expressed by IPTG induct ion. The protein was about 46 ku. Western blot showed specific react ion of the expressed product with type A FMDV ant ibody- posit ive serum, and the protein had nice reaction activity.

Key words: FMDV, 3D polymerase gene, Prokaryotic expression

CLC Number:

Q945

ZHANG Yu, WANG Yong-lu, ZHANG Yong-guang, PAN Li. Cloning of 3D of Foot-and-mouth Disease Virus Strain AF72 and Its Expression in Escherichia coli[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(6): 32-35. doi: 10.7668/hbnxb.2008.06.007.

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