禽源NME/NM23核苷二磷酸激酶2(NME2)基因的克隆及表达鉴定  

Cloning,expression and identification of the Gallus NME/NM23 nucleoside diphosphate kinase 2 gene from chicken

作　　者：谢丽基 谢芝勋 王盛 黄娇玲 邓显文 谢志勤 罗思思 曾婷婷 张艳芳 张民秀 范晴 XIE Liji;XIE Zhixun;WANG Sheng;HUANG Jiaoling;DENG Xianwen;XIE Zhiqin;LUO Sisi;ZENG Tingling;ZHANG Yanfang;ZHANG Minxiu;FAN Qing(Guangxi Veterinary Research Institute/Guangxi Key Laboratory of Veterinary Biotechnology,Nanning 530001,China)

机构地区：[1]广西壮族自治区兽医研究所/广西兽医生物技术重点实验室,广西南宁530001

出　　处：《畜牧与兽医》2021年第4期113-116,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31660715);广西科技专项(AA17204057,AD17195083);“广西八桂学者”专项(2019-79);国家“万人计划”领军人才专项(W02060083)。

摘　　要：参照GenBank中禽源核苷二磷酸激酶2(NME2)基因序列,设计了NME2基因的特异性引物。采用RT-PCR扩增NME2基因,经双酶切后克隆到真核表达载体pEF1α-Myc,得到重组质粒pEF1α-Myc-NME2。将构建好的重组质粒pEF1α-Myc-NME2转染DF1细胞后,采用间接免疫荧光和Western blot技术对目的蛋白进行验证。结果表明,Myc-NME2融合蛋白在DF1细胞内得到了正确表达。本研究为后续研究禽源NME2基因的功能奠定了基础。According to the sequences of the Gallus NME/NM23 nucleoside diphosphate kinase 2 NME2 gene in GenBank,one pair of primers was designed and synthesized.The NME2 gene from chicken was amplified by reverse transcription chain reaction(RT-PCR),and was inserted into the plasmid pEF1α-Myc vector.The recombinant plasmid containing the NME2 gene was named pEF1α-Myc-NME2.The constructed eukaryotic expression recombinant plasmid was transfected into DF1 cells to express the NME2 proteins,and the expressed proteins were verified by indirect immunofluorescence and Western blot.The results showed that Myc-NME2 fusion proteins were correctly expressed in the DF1 cells.This study laid a foundation for further research on the function of the NME2 gene.

关 键 词：NME/NM23核苷二磷酸激酶2 克隆 表达 

分 类 号：S852.6[农业科学—基础兽医学]