慢性髓系白血病bcr-abl融合基因的克隆及其在COS-7细胞中的表达  

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

作　　者：刘楠[1] 孙秉中[1] 冯琦[1] 高杰英[2] 刘永全[2] 罗振革[2] 

机构地区：[1]第四军医大学西京医院血液科,陕西西安710032 [2]军事医学科学院微生物流行病研究所免疫室,北京100850

出　　处：《细胞与分子免疫学杂志》2000年第4期346-348,359,共4页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的克隆慢性髓系白血病(CML)融合基因bcr abl的cDNA序列 ,并在真核细胞中表达。方法以设计的两条引物扩增bcr abl融合位点两侧的cDNA ,测序后克隆至真核表达载体 pcDNA3.1 ,转染COS 7细胞。取常规培养的细胞进行RT PCR鉴定。结果①PCR扩增出490bp大小的cDNA ,其序列测定除第452位的碱基C突变为T外 ,其余序列均正确。②RT PCR法检测到转染细胞系中 ,有bcr ablmRNA的表达。结论成功地克隆了CML融合基因bcr abl融合位点两侧的490bp 序列并表达 ,为研制bcr abl基因疫苗治疗CML奠定了基础。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.

关 键 词：慢性髓系白血病 BCR-ABL融合基因 克隆 

分 类 号：R733.72[医药卫生—肿瘤]