Direct cloning of cDNA inserts from lambda gt11 phage DNA into a plasmid vector by a novel and simple method.
Bacteriophage lambda gt11 has been used quite extensively for producing cDNA libraries. The cDNA inserts are usually subcloned into a plasmid vector for large scale production and analysis. However, isolating the recombinant DNA of interest from the phage clones can be a tedious task. Since the E. coli strain Y1088 used for lambda gt11 phage infection carries a pBR322-derived plasmid endogenously, we reasoned that this endogenous plasmid could be used directly for cloning the cDNA phage insert. In this report, we describe a method in which cDNA inserts from lambda gt11 phage were cloned directly into the pBR322 plasmid vector, bypassing the time-consuming procedures of preparing plasmid DNA as a subcloning vector. This method is likely to be extended to the cloning of DNA inserts derived from other phage lambda vectors when bacteria containing endogenous pBR322 are used as host cells.
Chiu
I M
IM
Department of Internal Medicine, Ohio State University, Davis Medical Research Center, Columbus 43210.
Lehtoma
K
K
D010582:Bacteriophage lambda / Q000235:genetics* / Q000254:growth & development; D001921:Brain / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008025:Ligases; D010957:Plasmids
D004279:DNA, Viral; D008025:Ligases