粉尘螨Der f15的基因克隆与其表达载体的构建  

Cloning and Vectorconstuction of Der f15 from House Dust Mite Dermatophagoides Farina

作　　者：李钟鸣[1] 邬玉兰[1] 刘志刚[1] 

机构地区：[1]深圳大学医学院过敏反应与免疫学研究所,广东深圳518060

出　　处：《江西师范大学学报（自然科学版）》2013年第2期159-161,共3页Journal of Jiangxi Normal University(Natural Science Edition)

基　　金：国家"863"计划(2006AA02A231);国家自然科学基金(81071388);广东省自然科学基金(04554);广东省高等学校国际暨港澳台科技合作创新平台项目(2012gjhz0009);深圳市重点实验室组建项目(SW201110010);深圳市科技计划基础研究重点项目(JCYJ20120613100657482);深圳市南山区一般研发课题(KC2012JSYB0003A)资助项目

摘　　要：先挑取纯培养的粉尘螨,提取总的RNA,后采用RT-PCR方法进行反转录出cDNA.由cDNA提取出目的基因进行片段扩增,产物连接入T载体(pMD18-T).经扩增后,用EcoR I和Xho I双酶切,将目的基因分别连接到pET28和pET32的表达载体上,得到重组质粒pET28和pET32,即粉尘螨的基因克隆与表达载体构建成功.The living mites of south China area which had been identified and cultured were picked, from which extractedthe total RNA. RT-PCR technology were used to clone the gene and then connected it to the T vector （pMD18-T）. After that,the expression vector （pET28 and pET32） was constructed for the new gene segment and were connect ed together.

关 键 词：粉尘螨 Der f15 克隆 载体构建 

分 类 号：Q78[生物学—分子生物学]