Expression and Identification of Porcine Antimicrobial Peptide PBD-1 in Pichia pastoris |
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| Authors: | JIANG Xue-bin CHEN Jia-wei YANG Jun LIU Shun-zhi XIAO An-ji LU Zhi-peng CHU Pin-pin HUANG Chao-yuan CAI Hai-ming MA Miao-peng ZHANG Ling-hua |
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| Affiliation: | 1. Research Institute of Bioengineering of Guangzhou University, Guangzhou 510006, China;2. Function and Regulation of Agricultural Biotechnology Protein of Guangdong Province Key Laboratory, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | In order to produce antimicrobial peptide PBD-1 with bioactivity in Pichia pastoris,according to published amino acid sequence of PBD-1 and the partiality codon of yeast,the PBD-1 gene was amplified by SOE-PCR and cloned into pPIC9K to construct a recombinant expression vector pPIC9K-PBD-1.The recombinant vector was linearized by SacⅠ,and then transformed into SMD1168 by electroporation.Positive yeast expression strain was obtained by PCR.The antimicrobial peptide PBD-1 (approximately 4.5 ku) was expressed by methanol induction.Antibacterial activity assay showed that the antimicrobial peptide PBD-1 had better antibacterial activity against E.coli,S.aureus and B.subtilis. |
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| Keywords: | PBD-1 gene expression Pichia pastoris PCR |
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