CONSTRUCTION OF PLASMID CLONING VECTORS FOR LACTIC STREPTOCOCCI WHICH ALSO REPLICATE IN BACILLUS-SUBTILIS AND ESCHERICHIA-COLI
CONSTRUCTION OF PLASMID CLONING VECTORS FOR LACTIC STREPTOCOCCI WHICH ALSO REPLICATE IN BACILLUS-SUBTILIS AND ESCHERICHIA-COLI
复制标题
DOI：
10.1128/aem.48.4.726-731.1984
复制
发表时间：
1984-01-01
影响因子：
4.4
通讯作者：
VENEMA, G
中科院分区：
文献类型：
--
作者：
KOK, J;VANDERVOSSEN, JMBM;VENEMA, G
神秘的乳脂链球菌 Wg2 质粒 pWV01（1.5 兆道尔顿）带有来自 pC194 的氯霉素抗性 (Cmr) 基因。重组质粒（pGK1，2.4兆道尔顿）在枯草芽孢杆菌中复制并表达Cmr。通过插入来自 pE194 cop-6 的红霉素抗性 (Emr) 基因，从该质粒构建插入失活载体。该质粒（pGK12，2.9兆道尔顿）在Emr基因中包含独特的BclI位点，在两个抗性基因之外包含独特的ClaI和HpaII位点。它以大约.apprx的拷贝数稳定地维持在枯草芽孢杆菌中。 5. pGK 12 还将大肠杆菌感受态细胞转化为 Cmr 和 Emr。大肠杆菌中的拷贝数为.apprx。 60.pGK12转化的乳酸链球菌原生质体。在该宿主中，两种抗性基因均表达。 pGK12 在乳酸链球菌中稳定维持在 3 拷贝数。
The cryptic Streptococcus cremoris Wg2 plasmid pWV01 (1.5 megadaltons) was genetically marked with the chloramphenicol resistance (Cmr) gene from pC194. The recombinant plasmid (pGK1, 2.4 megadaltons) replicated and expressed Cmr in B. subtilis. From this plasmid an insertion-inactivation vector was constructed by inserting the erythromycin resistance (Emr) gene from pE194 cop-6. This plasmid (pGK12, 2.9 megadaltons) contained a unique BclI site in the Emr gene and unique ClaI and HpaII sites outside both resistance genes. It was stably maintained in B. subtilis at a copy number of .apprx. 5. pGK 12 also transformed E. coli competent cells to Cmr and and Emr. The copy number in E. coli was .apprx. 60. pGK12 transformed protoplasts of S. lactis. In this host, both resistance genes are expressed. pGK12 is stably maintained in S. lactis at a copy number of 3.