多头带绦虫蛋白激酶A催化亚基基因的克隆及原核表达

doi:10.16431/j.cnki.1671-7236.2017.10.005

Abstract

Abstract:

To study the potential of protein kinase A catalytic subunit of Taenia multiceps (TmPKA-C) gene as a diagnostic antigen, the TmPKA-C gene was amplified by RT-PCR from the total RNA of Taenia multiceps. The TmPKA-C gene fragment was ligated into prokaryotic expression vector pET-30a and transformed into Escherichia coli Transetta (DE3) strain, then the reactonogenicity of recombinant TmPKA-C protein was analyzed by Western blotting and indirect ELISA. The open reading frame of TmPKA-C gene was 1 032 bp, encoding 343 amino acids. The expression product of TmPKA-C gene mainly existed in the form of inclusion body, and the molecular weight of recombinant protein was about 42 ku. Western blotting analysis showed that the recombinant protein could react specifically with the sera from naturally infected sheep with coenurosis, and it could also have specific reaction with the sera of artificial infected sheep with coenurosis. The indirect ELISA analysis showed that the sera from the coenurosis of sheep could react specifically with the recombinant protein. These results proved that the recombinant protein had good reactionogenicity. This study laid a foundation for the search of new diagnostic antigens for coenurosis of sheep.

Key words: Taenia multiceps; TmPKA-C; prokaryotic expression; reactinogenicity

CLC Number:

S852.73⁺4

YANG Yang, LI Wen-hui, ZHANG Nian-zhang, LI Ting-ting, LI Li, YAN Hong-bin, JIA Wan-zhong, FU Bao-quan. Cloning and Prokaryotic Expression of Protein Kinase A Catalytic Subunit Gene from Taenia multiceps[J]. , 2017, 44(10): 2858-2864.

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Cloning and Prokaryotic Expression of Protein Kinase A Catalytic Subunit Gene from Taenia multiceps

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