LYRM1基因绿色荧光蛋白融合载体的构建及表达  

Construction and expression of green fluorescent protein fusion vector of LYRM1

作　　者：邱洁[1] 郭锡熔[2] 周晓玉[1] 程锐[1] 曹兴国 王玢[2] 张敏[2] 

机构地区：[1]南京医科大学附属南京儿童医院新生儿科,江苏南京210008 [2]南京医科大学附属南京妇幼保健院儿科,江苏南京210004

出　　处：《中国儿童保健杂志》2009年第6期667-669,共3页Chinese Journal of Child Health Care

基　　金：国家自然科学基金(30801256);江苏省自然科学基金(BK20080778)

摘　　要：【目的】构建LYRM1基因绿色荧光蛋白融合载体,进而观察LYRM1基因编码蛋白在细胞内的定位情况。【方法】运用RT-PCR技术从人网膜脂肪组织中分离LYRM1基因的完整编码框,将其亚克隆到绿色荧光蛋白表达载体pEGFP-N2,脂质体转染3T3-L1前体脂肪细胞,激光共聚焦显微镜下观察融合蛋白的表达情况。【结果】PCR、酶切鉴定及测序结果表明重组质粒构建正确。激光共聚焦显微镜下观察到pEGFP-N2转染的细胞中绿色荧光均匀分布于整个细胞,而pEGFP-LYRM1转染的细胞中绿色荧光主要集中在细胞核区域。【结论】成功构建了LYRM1绿色荧光蛋白融合载体,LYRM1编码蛋白在细胞中定位于胞核中。[Objective] To construct the green fluorescent protein fusion vector of LYRM1 and investigate the sub cellular localization of LYRM1. [Methods] The complete coding sequence of LYRM1 gene was amplified by RT-PCR from human omental adipose tissue and was subcloned into pEGFP-N2 vector. The recombinant plasmid was then trans feeted into 3T3-L1 preadipocytes by liposome. The subcellular localization of LYRM1 gene was observed by laser scanning confocal microscopy. [Results] The recombinant plasmid was confirmed by PCR, restriction enzyme digestion and sequencing. The green fluorescence protein was distributed in the whole cell transfected with pEGFP N2, but mainly distributed in nucleus transfected with pEGFP-LYRM1. [Conclusions] The green fluorescent protein fusion vector of LYRM1 is constructed successfully and expressed primarily in nucleus.

关 键 词：肥胖 LYRM1基因 绿色荧光蛋白 亚细胞定位 

分 类 号：R153.2[医药卫生—营养与食品卫生学]