水牛转录因子Sox2和Lin28克隆与表达研究

Cloning and Expression of Buffalo Transcription Factors Sox2 and Lin28

本研究克隆水牛特异性转录因子Sox2和Lin28,构建表达载体,为进一步研究2个基因在水牛干细胞多能性调控中的作用奠定基础。以胎牛生殖嵴总RNA为模版,采用RT-PCR扩增Sox2和Lin28的编码区序列(CDS),构建逆转录表达载体pMSCV-Sox2和pMSCV-Lin28,转入NIH3T3细胞,鉴定其表达情况。结果表明:水牛Sox2和Lin28的CDS全长分别为966 bp和618 bp,蛋白结构保守;Sox2、Lin28氨基酸序列与牛、猪、人和鼠相应氨基酸序列的相似性分别为96%、95%、94%、94%和98%、97%、89%和91%;pMSCV介导的Sox2和Lin28能在NIH3T3细胞中表达。

Buffalo transcription factors Sox2 and Lin28 were cloned and expressing vectors were constructed. This investigation will provide a good foundation for researching the roles of two genes in pluripotent regulation of buffalo stem cells. The that RNA was extracted from fetus germ ridge, the coding sequences （CDS） of Sox2 and Lin28 were cloned by RT-PCR. The expression retroviral vectors pMSCV-Sox2 and pMSCV-Lin28 were constructed, and transfected into NIH3T3 cells with retrovirus infection, expression of Sox2 and Lin28 were detected by RT-PCR respectively. The results showed that the CDS length of Sox2 and Lin28 were 966 bp and 618bp respectively. The protein structure exhibited high conserved, amino acids sequence exhibited high homology with bovine, pig, human and mouse, the percentage of similarity were 96%, 95%, 94%, 94% respectively for Sox2, and 98%, 97%, 89%, 91% for Lin28. pMSCV retrovirat vectors based Sox2 and Lin28 could expressed in the NIH3T3 cells respectively.