Cloning and expression of chronic myelogenous leukemia bcr-abl fusion gene in COS-7 cells

Aim Chronic myelogenous leukemia(CML) bcr abl fusion gene cDNA was cloned and expressed in COS 7 cells. Methods Two primers were designed for the amplification of bcr abl gene segment located around the fusion site by PCR. After the PCR product was confirmed by the sequence analysis, and then it was properly inserted into the expression vector pcDNA3.1,which was transfected in COS 7 cells. Transfected COS 7 cells were identified by RT PCR after routine cultivation. Results ①bcr abl fusion gene of 490 bp was obtained by PCR,and sequence analysis showed that only base C in position 452 mutated into T. ②RT PCR confirmed that bcr abl had been expressed in transfected COS 7 cells. Conclusion A bcr abl fusion gene of 490 bp is cloned and expressed successfully. This study lay the foundation for gene vaccine therapy of CML.