An expeditious method for constructing T-vectors usingEam 1105 I cassettes

Abstract

An expeditious method is described for constructing T-vectors containing complementary 3′-thymidine overhangs. A T-vector was developed by cloning a 90-bpEam 1105 I cassette containing 2Eam 1105 I restriction sites into a modified pUC119 vector. TheEam 1105 I cassette was generated by PCR with 2 specific primers containing different recognition sequences ofEam 1105 I. The recombinant vector was easily converted into a T-vector by digestion of the plasmid withEam 1105 I. The cloning efficiency of the PCR product was approximately 90%. The method described here is a simple way to construct a variety of T-vectors.