蛋白转导结构域-bcr/abl融合基因的构建和表达  

Construction and expression of a vector containing protein transduction domain and bcr/abl fusion gene

作　　者：梁英民[1] 孙强[2] 蒋姗姗[1] 王冀姝[2] 吴绒丽[1] 陈萍[2] 刘利[1] 韩骅[2] 

机构地区：[1]第四军医大学唐都医院血液科,西安710038 [2]第四军医大学遗传与发育生物学教研室

出　　处：《中华血液学杂志》2002年第1期5-8,共4页Chinese Journal of Hematology

摘　　要：目的　构建含蛋白转导结构域 (PTD)与慢性粒细胞白血病 (CML)bcr/abl融合基因片段的质粒 ,并在大肠杆菌中表达。方法　PCR扩增的CMLbcr/abl基因片段 ,经DNA测序后 ,与合成编码PTD的DNA片段一起插入质粒pET 16b ,构建表达载体pEPb ,转化大肠杆菌并进行了PTD bcr/abl蛋白的诱导表达和纯化。结果　跨越bcr/abl断裂点的 5 2 3bp的目的片段被有效地扩增。DNA序列分析表明所构建的含PTD bcr/abl融合基因的质粒与设计相同。PTD bcr/abl融合蛋白在转化大肠杆菌获得了高效表达并纯化。结论　成功地获得了PTD bcr/abl融合蛋白片段的基因表达产物 。Objective To construct a vector containing protein transduction domain (PTD) and bcr/abl fusion gene of chronic myelogenous leukemia and express PTD bcr/abl fusion protein in E. Coli. Methods DNA fragment encoding PTD was synthesized and fused to PCR amplified bcr/abl gene fragment, then inserted into plasmid pET 16b to get the expression vector pEPb containing PTD bcr/abl fusion gene, which was transfected and expressed in E. Coli LB21. PTD bcr/abl fusion protein was purified by affinity chromatography. Results 523 bp bcr/abl fusion gene was effectively amplified. The PTD bcr/abl gene sequencing showed the same sequence as scheduled. The fusion peptide was successfully expressed in E. Coli and purified. Conclusion The results may provide a new PTD bcr/abl fusion peptide for the immunotherapy of CML.

关 键 词：遗传转导 BCR/ABL融合基因 白血病 慢性 基因表达 大肠杆菌 

分 类 号：R733.72[医药卫生—肿瘤]