| 大黄鱼β_2-微球蛋白基因的克隆及其在大肠杆菌中的重组表达; Cloning of β_2-microglobulin in large yellow croaker(Pseudosciaena croces) and efficient expression in Escherichia coli | |
| 余素红 ; 王玉桥 ; 陈新华 | |
| 2009 | |
| 关键词 | 大黄鱼 β2-微球蛋白 克隆 重组表达 Western-blotting large yellow croaker β2-microglobulin cloning recombinant expression Western-blotting |
| 英文摘要 | 主要组织相容性复合体(MAJOr HISTOCOMPATIbIlITy COMPlEX,MHC)是一类高度多态的基因群,它广泛分布于各种脊椎动物体内,在启动特异性免疫应答中起关键作用.MHC根据结构分为I型和II型两种.β2-微球蛋白(β2-MICrOglObulIn,β2M)是MHCI型分子的轻链.本文根据大黄鱼β2M的表达序列标签(EXPrESSEd SEQuEnCE TAgS,EST)序列设计合成了特异性引物,利用rT-PCr技术从大黄鱼脾脏组织中克隆了β2M基因(PSCr-β2M);构建了含β2M基因的原核表达载体(PgEX4T-2-β2M),在0.1MMOl/dM3IPTg诱导下重组β2M得到了表达;采用SEPHArOSE 4b亲合层析纯化目的蛋白并制备了抗体;WESTErn-blOTTIng分析证实了该抗体具有与天然大黄鱼脾脏组织中β2M蛋白反应的特性.这些结果为进一步研究鱼类β2M结构与功能的关系,以及制备MHCI类分子四聚体奠定基础.; Major hiscompatibility complex(MHC) including MHC class I and MHC class II is an extended cluster of genes with extraordinary polymorphism spreading over nearly all vertebrates extensively and plays a critical role in the induction of immune responses.β2-microglobulin(β2m) is a light chain of MHC class I molecule.In this experiment,the β2m gene was cloned from spleen tissue of large yellow croaker by expressed sequence tags(EST) analysis and RT-PCR techniques.The expression vector pGEX-4T-2-β2m gene was presented,in which the mature peptide sequence of β2m gene was inserted.High-yield expression of β2m was achieved induced by 0.1mmol/dm3 IPTG in E.coli transformed with the expression vector,and protein was completely purified using glutathione sepharose 4B affinity chromatography methods for preparing specific antibody.Western blotting assay indicated that the antibody against pGEX-4T-2-β2m could react specifically with the native Pscr-β2m protein in spleen tissue.The high quality Pscr-β2m laid a good foundation for preparation of MHC I tetramer and the study on the relationship between the configuration of fish MHC I and its functions.; 国家自然科学基金资助项目(4047605230571439) |
| 语种 | zh_CN |
| 内容类型 | 期刊论文 |
| 源URL | [http://dspace.xmu.edu.cn/handle/2288/118860]  |
| 专题 | 生命科学－已发表论文 |
| 推荐引用方式 GB/T 7714 | 余素红,王玉桥,陈新华. 大黄鱼β_2-微球蛋白基因的克隆及其在大肠杆菌中的重组表达, Cloning of β_2-microglobulin in large yellow croaker(Pseudosciaena croces) and efficient expression in Escherichia coli[J],2009. |
| APA | 余素红,王玉桥,&陈新华.(2009).大黄鱼β_2-微球蛋白基因的克隆及其在大肠杆菌中的重组表达.. |
| MLA | 余素红,et al."大黄鱼β_2-微球蛋白基因的克隆及其在大肠杆菌中的重组表达".(2009). |
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