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

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.