慢性髓性白血病bcr-abl融合基因的克隆与表达

CLONING AND EXPRESSION OF CML bcr-abl FUSION GENE FRAGMENT

以慢性髓性白血病K562细胞总RNA为模板,采用RT PCR技术扩增包含bcr abl融合位点周围的基因片段,定向克隆到pGEX 6P 1载体谷光甘肽S转移酶(GST)的下游。将重组质粒转化大肠杆菌BL21菌株,以1.0mmol·L-1IPTG诱导bcr abl融合基因片段的表达,其产物经聚丙烯酰胺凝胶电泳鉴定,结果证实该融合蛋白分子质量约42ku。用该表达产物免疫ICR小鼠,所制备的抗血清可与K562细胞中特异性抗原反应,表明该bcr abl/GST融合蛋白具有bcr abl基因产物的特异抗原性。

The 450 bp of bcr\|abl fusion gene was amplified from total RNA of K562 cells by RT\|PCR and was cloned into the downstream of GST gene in pGEX\|6P\|1 vector. The recombinant vector was transformed into E.coli BL21 cell for expression. The size of fusion protein was 42 ku which identified by SDS\|PAGE. The ICR mice were immunized with the fusion protein. The anti\|serum could bind P210bcr\|abl in K562 cells in IFA, this results suggested that the fusion protein had specific antigenicity of P210bcr\|abl.