小鼠β2m正义与反义RNA表达载体的构建及鉴定  

Construction and Identification of the Mouse β2m Sense and Antisense RNA

作　　者：孟瑛[1] 李树浓[1] 余新炳[2] 黄绍良[3] 徐劲[2] 吴忠道[2] 

机构地区：[1]中山医科大学病理生理教研室,广东广州510089 [2]中山医科大学寄生虫学教研室,广东广州510089 [3]中山医科大学孙逸仙纪念医院儿科,广东广州510089

出　　处：《中山医科大学学报》2001年第6期414-417,共4页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：国家自然科学基金资助项目 ( 3980 0 134);广东省青年科学基金资助项目 ( 980 6 81);中山医科大学"2 11工程"重点课题基金资助项目 ( 2 0 110 2 -110 5 )

摘　　要：【目的】构建小鼠 β2微球蛋白基因 (β2microglobulingene,β2m)正义及反义RNA表达载体 ,为研究降低细胞MHCⅠ类分子的表达作准备。【方法】从小鼠基因组获得的 β2m克隆中以 β2m pSV2ΔHXgpt为模板 ,合成两对引物 ,采用常规PCR方法分别扩增出 β2m引导区及其下游主要编码区 ,即外显子 1及部分外显子 2 ,然后利用重叠延伸法将两条片段拼接起来 ,再将其分别正向、反向插入载体 pcDNA3 ,并进行限制性内切酶酶切鉴定及序列分析。【结果】构建出小鼠 β2m正义和反义RNA表达载体 pcDNA3 β2mSN及 pcDNA3 β2mAN。【结论】克隆的正义及反义 β2m经限制性内切酶酶切鉴定结果正确 ,测序分析结果与文献一致 ,从而为下一步降低移植排斥反应的体内外研究奠定了基础。To construct the mouse β2 microglobulin gene (β2m) sense and antisense RNA into expresion vectors, for further study on downregulating the expression of MHCⅠmolecules. The leader segment and the main coding blocks, exon 1 and part of exon 2, were amplified respectively by PCR from the β2m clone β2m pSV2ΔHXgpt. They were spliced together by overlap extension, then digested and inserted in sense or antisense orientation into plasmid pcDNA3. The two recombinant plasmids were identified by restriction endonuclease and the inserting genes were sequenced. The recombinant plasmids of the β2m sense and antisense RNA, pcDNA3 β2mSN and pcDNA3 β2mAN were obtained. [Conclusion] pcDNA3 β2mSN and pcDNA3 β2mAN cloning vectors are successfully constructed, and the β2m sequence is identical with that of the previous reports in literaure.

关 键 词：基因 MHC I类 Β2微球蛋白 小鼠 载体 动物实验 鉴定 抗原 

分 类 号：R392.11[医药卫生—免疫学]