清道夫受体蛋白的原核表达及初步鉴定  

Prokaryotic expression and preliminary identification of scavenger receptor protein

作　　者：许甜甜[1] 朱冰[1] 王长兵[1] 钟家禹[1] 林正方 

机构地区：[1]广州医科大学附属广州市妇女儿童医疗中心中心实验室,广东广州510120

出　　处：《国际检验医学杂志》2014年第12期1523-1525,共3页International Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目(309726300)

摘　　要：目的表达清道夫受体B2蛋白(SCARB2),初步鉴定表达产物。方法培养RD细胞,提取其mRNA经逆转录PCR(RT-PCR)方法扩增出scarb2基因片段,构建表达载体pET28a(+)/SCARB2,在原核表达系统(大肠杆菌BL21)中表达SCARB2蛋白,并用His单抗对表达蛋白进行免疫印迹法(Western blot)鉴定。结果构建的重组质粒pET28a(+)/SCARB2经异丙基-β-D-硫代半乳糖苷(IPTG)诱导,重组蛋白SCARB2高效表达并纯化成功,经His单抗初步验证有目的蛋白表达。结论表达并初步鉴定了SCARB2蛋白,为肠道病毒71型(EV71)与清道夫受体蛋白的作用机制及清道夫受体的单克隆制备提供参考。Objective To express and preliminary identify scavenger receptor B2 （SCARB2）protein.Methods SCARB2 cDNA was amplified by reverse transcription-polymerase chain reaction （RT-PCR）from mRNA extracted from RD cells,and cloned into pMD19-T vector.Expression vector pET28a（＋）/SCARB2 was constructed,and SCARB2 protein was expressed in prokaryotic ex-pression system.Obtained recombinant proteins were identified by detection of Western blot using His labeled monoclonal antibody.Results Recombinant SCARB2 proteins,expressed by induction of isopropy-β-D-thiogalactoside （IPTG）,were success-fully purified and specifically recognized by His labeled monoclonal antibody.Conclusion SCARB2 proteins could be expressed and identified successfully,which could provide reference for researching mechanism of enterovirus type 71 （EV71）and scavenger re-ceptor protein,and for preparation of SCARB2 monoclonal antibodies.

关 键 词：肠道病毒71 型 清道夫受体 基因克隆 原核表达 

分 类 号：R725.1[医药卫生—儿科]