携带噬菌体λ cI基因的质粒构建
Construction of plasmids carrying the cI gene of bacteriophage lambda.
作者信息
Backman K, Ptashne M, Gilbert W
出版信息
Proc Natl Acad Sci U S A. 1976 Nov;73(11):4174-8. doi: 10.1073/pnas.73.11.4174.
Abstract
By techniques of recombination in vitro, we have constructed a plasmid bearing the repressor gene (cI) of bacteriophage lambda fused to the promoter of the lac operon. Strains carrying this plasmid overproduce lambda repressor. This functional cI gene was reconstituted by joining DNA fragments bearing different parts of that gene. Flush end fusion techniques, involving no sequence overlap, were necessary for the construction; in certain cases, the abutting of the DNA molecules bearing ends generated by different restriction endonucleases creates a sequence at the junction which is recognized by one of the restriction endonucleases.
摘要
通过体外重组技术，我们构建了一个质粒，该质粒带有与乳糖操纵子启动子融合的噬菌体λ阻遏基因（cI）。携带此质粒的菌株会过量产生λ阻遏物。这个功能性的cI基因是通过连接携带该基因不同部分的DNA片段而重建的。构建过程需要采用不涉及序列重叠的平端融合技术；在某些情况下，由不同限制性内切核酸酶产生末端的DNA分子邻接会在连接处产生一个能被其中一种限制性内切核酸酶识别的序列。
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