乳链菌肽前体基因(nisZ)在乳酸乳球菌中的克隆和表达  

Cloning and Expression of nisZ Gene in Lactococcus lactis

作　　者：陈秀珠[1] 胡海菁[1] 杨巍[1] 还连栋[1] 

机构地区：[1]中国科学院微生物研究所微生物资源前期开发国家重点实验室,北京100080

出　　处：《Acta Genetica Sinica》2001年第3期285-290,共6页

基　　金：国家自然科学基金!(3957008);国家"九五"科技攻关!(96-C3-01-5);中国科学院重点研究项目&&

摘　　要：用ＰＣＲ技术从克隆有完整乳链菌肽生物合成基因簇（来自于乳链菌肽高产菌株Ｌ． ｌａｃｔｉｓ ＡＬ２）的重组噬菌体λＨＪ－３中扩增了编码乳链菌肽的前体基因，与ｐＭＧ３６ｅ连接得到重组质粒ｐＨＪ２０１，用电击转化法将ｐＨＪ２０１转化到Ｌ ｌａｃｔｉｓ ＮＺ９８００中，经活性测定和Ｔｒｉｃｉｎｅ－ＳＤＳ－ＰＡＧＥ电泳证实乳链菌肽前体基因获得了功能表达．ＤＮＡ序列分析表明乳链菌肽高产菌株Ｌ ｌａｃｔｉｓ ＡＬ２产生的是ＮｉｓｉｎＺ．发现ｐＨＪ２０１在Ｌ ｌａｃｔｉｓ ＮＺ９８００中有良好的稳定性。The gene encoding the precursor of nisin was amplified by PCR using the HJ-3 DNA as the template, which contained the entire nisin biosynthesis gene cluster from Lactococcus lactis AL2 with high yield of nisin, and was cloned into pMG36e. The recombinant plasmid pHJ201 was introduced into Lactococcus lactis NZ9800 by electroporation. pHJ201 is very stable in L lactis NZ9800. Antimicrobial activity test and Tricine-SDS-PAGE analysis revealed that L lactis NZ9800 harbouring pHJ201 restored ability of nisin production, but the production level was markedly lower than L lactis AL2. The result of DNA sequence analysis indicated that Nisin Z is produced by L lactis AL2.

关 键 词：乳酸乳球菌 乳链菌肽前体基因 基因克隆 表达 

分 类 号：Q786[生物学—分子生物学]