IN-VIVO CLONING OF PCR PRODUCTS IN ESCHERICHIA-COLI
IN-VIVO CLONING OF PCR PRODUCTS IN ESCHERICHIA-COLI
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DOI：
10.1093/nar/21.22.5192
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发表时间：
1993-11-11
影响因子：
14.9
通讯作者：
VOGELSTEIN, B
中科院分区：
文献类型：
--
作者：
OLINER, JD;KINZLER, KW;VOGELSTEIN, B
本文介绍了一种利用内源性大肠杆菌酶活性克隆PCR产物的有效方法。PCR产物被设计成包含与线性化载体两端序列相同的末端序列。然后将PCR产物和载体DNA简单地共转染到大肠杆菌菌株JC8679中，无需对PCR产物进行酶处理或体外连接。该菌株的高同源重组率导致插入物有效地整合到载体中，我们称之为体内克隆（IVC）的过程。
This report describes an efficient method to clone PCR products exploiting endogenous Escherichia coli enzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. The high rate of homologous recombination in this strain results in efficient incorporation of the insert into the vector, a process we refer to as in vivo cloning (IVC).