Molecular cloning and eukaryotic expression of chronic myelogenous leukemia bcr-abl fusion gene

Objective To provide the target gene and target antigen for the development of new vaccine against chronic myelogenous leukemia(CML).Methods According to the gene sequence encoding bcr abl(b3a2)fusion gene from CML K562 cell line,we designed a pair of oligonucleotide primers,obtained the 472bp gene fragment of bcr abl by using reverse transcription polymerase chain reaction(RT PCR),and inserted the gene into plasmid pcDNA3 1 via pGEM T easy vector to construct recombinant plasmid.The recombinant plasmid was transfected into CHO cell.The expression product of the gene fragment was analyzed by using indirect fluorescence immunoassay(IFA).Results The bcr abl fusion gene fragment encoding bcr abl of CML was amplified by using RT PCR.A recombinant eukaryotic expression plasmid was successfully constructed.The recombinant plasmid stably expressed the fragment of bcr abl fusion gene in CHO cell cytoplasm transfected with liposome through IFA.Conclusion Bcr abl fusion gene was expressed in CHO cells.The results of this investigation provided a basis for further research on the development of bcr abl fusion gene vaccine of CML.