【摘要】 目的 :克隆和表达Bcr abl融合基因融合位点周围的基因片段。方法 :以慢性粒细胞白血病 (CML)K5 6 2细胞株总RNA作为模板 ,采用RT PCR方法扩增包含Bcr abl(b3a2 )融合位点周围的基因片段。将RT PCR产物按正确的阅读框架 ,定向克隆进pGEX 6P 1载体谷胱甘肽 S 转移酶 (GST)的下游 ,将重组质粒转化大肠杆菌BL2 1菌株 ,以 1.0mmol/LIPTG诱导Bcr abl融合基因片段的表达。结果 :原核细胞融合表达产物经聚丙烯酰胺凝胶电泳鉴定 ,证实该融合蛋白为 4 2 0 0 0大小。用该表达产物免疫ICR小鼠 ,所制备的抗血清可与K5 6 2细胞特异性结合。结论 :原核细胞表达的p4 2 Bcr abl融合蛋白具有p2 10 Bcr abl融合蛋白的特异抗原性 ,为进一步实验研究奠定了基础。更多还原

【Abstract】 Objective:To clone and express the fragment of Bcr abl fusion gene (b3a2).Method:The fragement of Bcr abl fusion gene was amplified from K562 cells of CML by RT PCR and was cloned into the downstream of GST gene in pGEX 6P 1 vector according to the current open reading frame.This recombinant vector was transformed into BL 21 bacteria for expression.Result:The 42 000 fusion protein was identified by SDS PAGE.We also used the 42 000 fusion protein to immunize the ICR mice for getting anti serum. The polyclone antibody was combined to K562 cells which are chronic myeloid leukemia through IFA.Conclusion:The fusion protein p42 Bcr abl GST has specific immunogenicity of p210 Bcr abl .更多还原