合浦珠母贝iPS细胞相关的转录因子的克隆及分析  

Cloning and sequence analysis of iPS related transcription factors from Pinctada fucata

作　　者：张若云[1] 徐广锐 潘聪[1] 谢莉萍[1] 王洪钟[1] 张贵友[1] 张荣庆[1] 

机构地区：[1]清华大学生命科学学院,北京100084

出　　处：《海洋科学》2014年第4期7-14,共8页Marine Sciences

基　　金：国家重点基础研究发展计划项目(2010CB126405)

摘　　要：转录因子Sox2、Oct4、c-Myc和KLF4在iPS细胞研究中具有重要作用,将它们加入体外培养的细胞中,可以诱导体细胞去分化成为多能干细胞。作者通过RACE-PCR技术从合浦珠母贝(Pinctada fucata)的外套膜组织中克隆获得了3个转录因子——c-Myc、Sox2及KLF4的cDNA。c-Myc全长1585bp,开放阅读框的长度为1131bp,编码376个氨基酸,蛋白结构分析表明它具有保守的HLH和Myc-N结构域。Sox2全长1908bp,开放阅读框长度为990bp,编码329个氨基酸,蛋白结构分析它具有保守的HMG-box和Soxp结构域。KLF4全长2268bp,开放阅读框长624bp,编码207个氨基酸,预测该蛋白具有两个zf-H2C2_2结构域。BLAST分析它们均与太平洋牡蛎的相关蛋白具有较高同源性,说明其与太平洋牡蛎亲缘关系更近。RT-PCR实验发现这3个基因在合浦珠母贝外套膜、足、生殖腺、内脏团、闭壳肌和鳃6种组织中均有表达,表明它们是非常保守的转录因子。本研究对于合浦珠母贝细胞系建立和研究具有启发意义。The four transcription factors named Sox2, Oct4, c-Myc and KLF4 can reprogram differentiated in vitro culture cells to an embryonic-like state, in another name, ips cells. The complete gene sequence of c-Myc, Sox2 and KLF4 from Pinctada fucata was amplified by RACE PCR. The complete sequence of c-Myc gene was 1585 bp and the complete ORF length was 1131 bp which encoded a protein with 376 amino acid residues. The analysis of pro-tein structure showed c-Myc protein had the conservative HLH and Myc-N domains, which have the DNA-binding function. Sox2 gene has the complete sequence of 1908 bp and the complete ORF length of 990 bp which encoded a protein with 329 amino acid residues. And the Sox2 protein had the conservative HMG-box and Soxp domains, which have the DNA-regulation function. KLF4 gene was 2268 bp long and the complete ORF length was 624 bp which encoded a protein with 207 amino acid residues. And the KLF4 protein had two zf-H2C2_2 domains, which have the DNA-binding function. Blast analysis indicated the three deduced amino acid sequences of the c-Myc, Sox2 and KLF4 genes from P. fucata showed high homology with proteins from Crassostrea gigas. RT-PCR result shows that these three genes were expressed in all six tissues from P. fucata including mantle, foot, gonal, visceral mass, adductor muscle and gill, suggesting that these genes were conservative transcriptional factors. Our study inspires the work on cell culture and research in P. fucata.

关 键 词：诱导多功能干细胞 转录因子 克隆 合浦珠母贝(Pinctada fucata) 

分 类 号：S823.3[农业科学—畜牧学]