杂合抗菌肽Sα-Jp基因真核表达载体的构建

摘要

To obtain the new hybrid antibacterial peptide of high efficient expression and antimicrobial activity. Ac-cording to the sequence of amino acids about Spinigerin α, Japonicin II antibacterial peptide, combined with the structure of the protein molecules and antibacterial mechanism of antibacterial peptides , choose pichia preference codon, design and synthesis hybrid antimicrobial peptide Sα-Jp gene, and directed cloning to eukaryotic expression vector pPICZαA. By polymerase chain reaction, the double enzyme digestion and the sequence analysis to identify the transformation of host insert is contained in the recombinant expression vector of pPICZαA-Sα-Jp. Results show that we have been successfully build a eukaryotic expression vector, and it can be used in the secretion of eu-karyotic expression.%为获取高效表达且具有抗菌活性的新型杂合抗菌抗菌肽。根据 Spinigerin α、Japonicin II 抗菌肽的氨基酸序列，结合蛋白质分子的结构和抗菌肽的抑菌机制，选用毕赤酵母偏爱密码子，设计合成杂合抗菌肽 Sα-Jp基因，并将其定向克隆至真核表达载体 pPICZαA 上。经聚合酶链式反应，双酶切，序列分析等鉴定，所转化的宿主菌中含有插入 Sα-Jp 基因的重组表达载体 pPICZαA-Sα-Jp。结果表明：已成功构建真核表达载体，可用于真核分泌表达。