非洲爪蟾基因MGC64236的克隆及初步分析  

Cloning and Preliminary Characterization of a Novel MGC64236 Gene in Xenopus laevis

作　　者：杨玲麟[1,2] 陈平[2] 胡火珍[2,3] 吴敬波[1] 

机构地区：[1]泸州医学院附属医院,四川泸州646000 [2]四川大学华西医院生物治疗国家重点实验室,成都610041 [3]四川大学生命科学学院,成都610064

出　　处：《四川动物》2010年第4期522-526,共5页Sichuan Journal of Zoology

基　　金：国家973计划(2004CB518800;2001CB510001)

摘　　要：MGC64236基因是本实验室用脐静脉内皮细胞免疫的兔血清筛选非洲爪蟾cDNA文库而鉴定的一个功能未知的基因。本研究提取非洲爪蟾受精卵总RNA通过RT-PCR得到基因MGC64236的开放读码框651bp、编码202个氨基酸;运用生物信息学研究工具进行分析,发现该基因编码的蛋白有3个潜在的跨膜域,有一保守的结构域DUF1370,可能通过其胞内部分的磷酸化机制在介导细胞内外的信号转导中发挥重要作用;在非洲爪蟾胚胎各个发育时期用RT-PCR检测该基因的表达情况,发现在非洲爪蟾胚胎发育的几个重要时期该基因都有高表达,而在成体则特异地表达于脑和眼等神经组织;构建绿色荧光融合蛋白真核表达载体并转染HEK293细胞,对MGC64236蛋白的亚细胞定位,发现MGC64236蛋白比较特异地表达在细胞膜。MGC64236 is an unknown gene that was identified in our laboratory through screening the Xenopus cDNA library with rabbit immune serum against human umbilical vein endothelial cells. In this study, the open reading frame of the MGC64236 gene was successfully cloned. Bioinformatics was applied to predict the potential functions of the MGC64236 protein. The expression profile of the MGC64236 gene in various tissues and developmental stages was also investigated. The recombinant vector expressing green fluorescence protein was constructed and transfected into HEK293 cells to identify the subcellular localization of the MGC64236 protein. The original data obtained through experiments and bioinformatics in this study provide a foundation for further functional studies of MGC64236.

关 键 词：非洲爪蟾 MGC64236 生物信息学 表达谱 亚细胞定位 

分 类 号：Q78[生物学—分子生物学]