冷诱导RNA结合蛋白CIRP过表达载体的构建-应用与环境生物学报.DOCVIP

冷诱导RNA结合蛋白（CIRP）过表达载体的构建及鉴定* 李静辉1收稿日期 Received 接受日期 Accepted*资助项目：国家自然科学基金项目（NO.；农业部948计划重点资助项目（NO. 2011-G35）；黑龙江省自然科学基金项目（NO. C201103）；研究生创新项目（NO.YJSCX2014-Y25）Supported by grant from the National Natural Science Foundation of China（No;“948”key program of the Ministry of Agriculture of China(No.2011-G35)；Project of Natural science foundation of Heilongjiang province of China(No. C201103)**通讯作者 收稿日期 Received 接受日期 Accepted *资助项目：国家自然科学基金项目（NO.；农业部948计划重点资助项目（NO. 2011-G35）；黑龙江省自然科学基金项目（NO. C201103）；研究生创新项目（NO.YJSCX2014-Y25）Supported by grant from the National Natural Science Foundation of China（No;“948”key program of the Ministry of Agriculture of China(No.2011-G35)；Project of Natural science foundation of Heilongjiang province of China(No. C201103) **通讯作者 Corresponding author (E-mail: lishize1@) 1.黑龙江八一农垦大学动物科技学院 ；2. 黑龙江八一农垦大学食品学院 大庆 163319 摘 要 构建携带冷诱导RNA结合蛋白(Cold inducible RNA-binding protein，CIRP)的过表达载体，为进一步研究 CIRP 的作用机制奠定良好的实验基础。首先从4 ℃条件下冷处理8 h的SD大鼠睾丸组织中获得 CIRP 基因的 cDNA 序列，然后扩增 CIRP 基因的编码区，将其克隆到真核表达载体pLenti6/V5中，酶切鉴定并测序。将克隆成功的质粒转染HEK293T细胞，用western blot 检测CIRP 的表达水平。本试验成功扩增了 CIRP 编码区，并将其克隆至载体 pLenti6/V5 中；当将CIRP 过表达载体转染 HEK293T 细胞后，western blot 检测结果显示 CIRP 蛋白表达水平明显增加(P0.01)。本研究成功构建了CIRP过表达载体，为今后的研究提供了基础工具。 图4表1参21 关键词 冷诱导RNA结合蛋白；过表达；质粒；冷应激 CLC Q494 : Q78 Construction and Identification of Over-expressing Vector of Cold inducible RNA-binding Protein (CIRP)* LI Jinghui1, MENG Yu1, LI Yuheng1, LI Changsheng1, YANG Huanmin1＆LI Shize1,2** 1.College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China 2.College of food, Heilongjiang Bayi Agricultural University, Daqing 163319, China Abstract Objective: To provide a basis tool for further research of CIRP function, a vector for the over-expression of Cold inducible RNA-binding Protein (CIRP) was constructed in this study. After obtained CIRP cDNA sequencing of testicular tissue from SD rats wi