具有真细菌基因启动子活性的盐生盐杆菌质粒DNA片段  

Plasmid DNA Fragments fro Halobacterium halobium Active as Eubacteria Promoters in Escherichia coli

作　　者：孙广秀[1] 江爱民[1] 沈萍[1] 

机构地区：[1]武汉大学生命科学学院

出　　处：《Acta Genetica Sinica》1997年第4期380-384,共5页

基　　金：国家自然科学基金

摘　　要：利用大肠杆菌启动子探测质粒ｐＫＫ２３２－８为载体、用两组限制性内切酶ＢａｍＨＩ－ＳａｌＩ和ＨｉｎｄⅢ－ＳａｌＩ分别消化盐生盐杆菌Ｊ７（Ｈａｌｏｂａｃｔｅｒｉｕｍｈａｌｏｂｉｕｍ）的质粒ｐＨＨ２０５，在体外进行重组，转化Ｅ．ｃｏｌｉＨＢ１０１感受态细胞，在含氨苄青霉素和氯霉素的选择平板上筛选转化子，并从随机挑选的２０株转化子中，获得抗氯霉素水平达到１１０μｇ／ｍｌ的转化子Ｔ１和Ｔ２，所含重组质粒分别被命名为ｐＪＨ和ｐＪＢ。经限制性酶切分析及杂交分析表明，ｐＪＨ质粒上插入了一段来源于ｐＨＨ２０５质粒的ＤＮＡ片段，其大小为８００ｂｐ左右。通过重新转化实验进一步表明，该ＤＮＡ片段在大肠杆菌中具有启动子功能，从而证明，在古细菌（盐生盐杆菌）的质粒ＤＮＡ中存在具有真细菌（大肠杆菌）基因启动子活性的ＤＮＡ片段。In this paper, a promoter-probe plasmid pKK232-8 was used as a vector which functioned inE. coli. The plasmid pHH205 of Halobacterium halobium J7 was digested by two groups of restriction endonucleases BamHI-SalI and HindⅢ-SalI, recombinated in vitro and transformed into E.coli. The transformants were selected on resistance plates to contain ampicillin (Am) and chloramphenicol (Cm). Fromrandom-selected 20 strains of transformants we obtained transformants T1and T2 , whose level of Cm resistance got to 110μg/ml .The recombinant plasmids fro T1 and T2 were named pJH and pJB, respectively. The resultes of analysis by restriction endonucleases digestion and molecular hybridization showed that recombinant plasmid pJH carried a inserted fragment with size of 800bp from plasmid pHH205. Experimental resultes of re-transfomation proved further that the DNA fragment had promoter funcion in E. coli. Thus. this indicatedthat DNA fragments from plasmid of archaebaeteria (H. halobium) may function as eubacteria (E. coil) promoters.

关 键 词：盐生盐杆菌 启动子活性 质粒DNA 真细菌基因 

分 类 号：Q939.103[生物学—微生物学]