摘要
利用巴斯德毕赤酵母(Pichia pastors)系统表达抗菌肽——牛乳铁多肽素(bovine lactoferricin,简称Lf-cinB),获得的分泌型表达产物具有较强的抗菌活性。首先将人工合成的LfcinB基因片段克隆到巴斯德毕赤酵母分泌型表达载体pPIC9K中,获得的重组质粒pPIC9K-LfcinB通过限制性内切酶SalⅠ酶切线性化,经电穿孔法转化入毕赤酵母细胞SMD1168内。G418抗性筛选,得到高拷贝转化子,经PCR检测LfcinB基因与毕赤酵母染色体稳定整合。阳性克隆经甲醇诱导表达LfcinB。结果表明,抗菌肽牛乳铁多肽素基因已经整合到酵母细胞基因组中并获得表达,表达产物具有较强的杀菌作用。
In present study, bovine Lactoferricin was first secretly expressed in Pichia pastors yeast expression system. The synthesized LfcinB gene fragment was cloned into expression vector pPIC9K, and then obtained recombinant plasmid, designated as pPIC9K-LfcinB, was linearized and transformed into Pichia pastors strains SMD1168 by electroporation. The transfonnants were screened with Geneticin and multiply-copy colonies were harvested, in which LfcinB gene was verified to inserted into yeast chromosome stably. The positive recombinant Pichia strains were induced with methanol to express LfcinB in culture supernatant. It＇s expressive products has high activity of killing bacteria. We concluded that LfcinB gene was cloned and integrated into yeast chromosomes, and obtained expression peptide was tested to have high antibacterial activity.
出处
《微生物学通报》
CAS
CSCD
北大核心
2007年第2期265-269,共5页
Microbiology China
