Expression Vector Construction and Antibacterial Activity Characterization of Caprine β-defensin

doi:10.11924/j.issn.1000-6850.2014-0969

Abstract

Abstract: This study was to construct the vector, shift in the Pichia pasroris eith high expression, and identify antibacterial activity characterization of the product, the results could help further research on the antibacterial mechanism. According to the CDS sequences for caprine β-defensin in GenBank, the author designed a pair of specific primers with the restriction enzyme cutting site to get the sequence, cloned it to the expression vector pPICZαA, contained the recombinant plasmid pPICZαA/GBD-1, and transformed into pichia pasroris by electroporation to induction expression. Sequence analysis showed that the amplicon 114 bp encoding a polypeptide of 38 amina acids, the molecular mass about 4.3 KD was expressed by using the Tricine-SDS-PAGE electrophoresis analysis, the recombinant protein had antibacterial activity and killed E.coli and Sta.aureus. The caprine β-defensin was successfully expressed in Pichia pastoris, which laid a foundation for further study of GBD-1 in the antibacterial mechanism and preparing the antibodies, and provide a theoretical basis for genetic engineering in the production of defensins.

Kang Jianfeng,Liu Guiqiong,Liu Shengmin and Jiang Xunping. Expression Vector Construction and Antibacterial Activity Characterization of Caprine β-defensin[J]. Chinese Agricultural Science Bulletin, 2015, 31(2): 40-44.

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