In vivo cloning of PCR products in E. coli.
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).
Oliner
J D
JD
Johns Hopkins Oncology Center, Baltimore, MD 21231.
Kinzler
K W
KW
Vogelstein
B
B
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D014169:Transformation, Bacterial
D009838:Oligodeoxyribonucleotides