乳酸乳球菌AL2基因文库的构建及乳链菌肽生物合成基因簇的筛选  

CONSTRUCTION OF GENOMIC LIBRARY FROM L.lactis AL2 AND ISOTATION OF ENTIRE NISIN BIOSYNTHESIS GENE CLUSTER

作　　者：陈秀珠[1] 胡海菁[1] 贾士芳[1] 陈美玲[1] 还连栋[1] 吴晓勇[2] 张成康 

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 [2]北京大学生命科学学院细胞生物学及遗传学系

出　　处：《微生物学报》2000年第5期465-469,共5页Acta Microbiologica Sinica

基　　金：国家自然科学基金资助项目! ( 3 9570 0 8) ;国家"九五"科技攻关项目!( 96 C0 3 0 1 0 5)&&

摘　　要：用本实验室获得的乳链菌肽高产菌株乳酸乳球菌AL2总DNA为供体 ,以λEMBL3为载体 ,构建了该菌株的基因文库 ,共获得 390 0多个噬斑。通过Southern杂交、PCR扩增及DNA序列测定 ,证实从该文库中筛选到一个含有完整乳链菌肽生物合成基因簇的重组噬菌体λHJ 3。A total DNA library of \%Lactococcus lactis\% AL2 with high yield of nisin was successfully constructed using lambda EMBL3 as vector. 3900 plaques were obtained, which is much more than the required number of recombinants that represents an entire \%L.lactis\% genome according to Clarke and Carbon formula. Southern hybridization, PCR amplification and DNA sequencing revealed that the entire nisin biosynthesis gene cluster was isolated from the constructed library.

关 键 词：乳酸乳球菌 基因文库 乳链菌肽 生物合成基因簇 

分 类 号：Q785[生物学—分子生物学]