Cloning and Fusion Expression of the gene encoding part of HTLV envelope protein gp21 / 中国输血杂志

ABSTRACT

Objective The gene encoding part of HTLV(Human Tcell leukemia viruses) envelope protein gp21 was cloned and expressed in order to prepare diagnostic agent for detecting HTLV.Methods The gene encoding part of HTLV envelope protein gp21 was obtained by combining four oligonucleotides into a long DNA fragment,then cloned and subcloned into Tvector and pGEX2T respectively.Expression was performed under induction in a fusion way that target protein gene was inserted downstream the carrying protein gene of pGEX2T which makes purification of target protein convenient.At last,Target protein was purified by affinity chromatography in glutathione sepharose 4B column and its antigenicity was confirmed by immunoblotting.Results and conclusion Sequence analysis showed it was feasible to obtain a target gene by ligating short oligonucleotides;As expected,target protein was expressed which exhibited strong antigenicity after being subcloned into fusion expression vector pGEX2T.