CRIF1基因的克隆表达  

Cloning and expression of CRIF1 gene

作　　者：冉茜[1] 单佑安[1] 李忠俊[1] 

机构地区：[1]第三军医大学附属新桥医院输血科,重庆400037

出　　处：《中国输血杂志》2010年第12期1018-1020,共3页Chinese Journal of Blood Transfusion

基　　金：国家科学基金资助项目(NO:30700336)

摘　　要：目的克隆表达CRIF1基因蛋白,为进一步研究该基因在骨髓微环境诱导白血病细胞G0/G1期阻滞过程中作用以及相关机制奠定基础。方法自HL60细胞中提取总RNA、逆转录,并与pET32a构建融合蛋白表达载体、蛋白原核表达载体;优化蛋白表达诱导条件,蛋白纯化;SDS-PAGE、Western Blot检测目的蛋白的表达。结果克隆并经测序获得全长744bp的CRIF1基因,成功构建融合蛋白表达载体和原核表达载体,优化并纯化蛋白;SDS-PAGE在42.5 kD处获得目的条带,Western Blot检测确定为CRIF1表达蛋白。结论成功克隆表达出CRIF1基因蛋白。Objective To clone and express CRIF1 gene in order to study the function and mechanism of CRIF1 gene,which could induce leukemic cells arrest in G0/G1 phase in the bone marrow microenvironment.Methods RNA was extracted from HL60 cells and transcribed reversely.CRIF1 gene was inserted into pET32a to construct fusion protein expression vector and prokaryotic expression vector.The condition for induction and expression was optimized.The fusion protein was purified,and detected by SDS-PAGE,and Western Blot.Results CRIF1 gene,of 774bp,was obtained and identified by sequencing.CRIF1 fusion protein expression vector and prokaryotic expression vector were constructed successfully.In optimal condition of induction and expression,highly purified protein was obtained.A protein band of about 42.5KD was detected by SDS-PAGE,and the protein was verified to be CRIF1 by Western Blot.Conclusion CRIF1 gene was cloned and expressed successfully.

关 键 词：细胞周期 CRIF1 白血病 

分 类 号：R457.1[医药卫生—治疗学] R557.1[医药卫生—临床医学]