摘要
以蓝细菌Ｐｌｅｃｔｃｎｅｍａｂｏｒｙａｎｕｍ内源小质粒ｐＰｂｓ（１．５ｋｂ）和Ｅ。ｃｏｌｉ质粒载体ｐＢＲ３２２（４．３ｋｂ）为材料，用限制性内切核酸酶ＣｌａⅠ分别消化，经Ｔ＿４ＤＮＡ连接酶连接，转化Ｅ．ｃｏｌｉＨＢ１０１感受态细胞，在含Ａｍｐ（Ａｐ）和含Ｔｅｔ（Ｔｃ）的ＬＢ琼脂培养基上筛选出Ａｍｐ￣ｒＴｅｔ￣ｓ的转化子，提取转化子的质粒，用ＣｌａⅠ消化，得１．５ｋｂ和４．３ｋｂ两片段，表明转化子所含质粒是ｐＰｂｓ和ｐＢＲ３２２的重组质粒，定名为ｐＰＲＳ－１。
Plasmid pPbs,endgenous plasmid of cyanobacterium plectonema boryanum,is 1.5 kb and has only one recognition site for Cla Ⅰ. Plasmid pBR322,vector plasmid of E. coli,is 4.3 kb and has Amp and Tet resistant genes, which are used as selective symbol genes. Therecognition site of Cla Ⅰis in the promoter of Tet resistant gene, If foreign DNA fragment is in-serted into this site, Tet resistant gene will lose its activity.Plasmids pPbs and pBR322 are di-gested by Cla Ⅰand are ligated by T_4 DNA ligase. The recombinant plasmids are transformed in-to E. coli HB101. Amp￣r and Tet￣s colonies are selected on LB agar midia, The plasmids in thesecolonies are digested into two fargments,1.5 kb and 4.3 kb, by Cla Ⅰ and one fragment of 5.8kb by Hind Ⅲ.It shows that this plasmid pPRS_1 is the recombinant plasmis of pPbs andpBR322.
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
1995年第2期256-260,共5页
Journal of Xiamen University：Natural Science
