Xu ZHENG, Jiao-fu YANG, Xiao-jing WANG, Yan LIANG, Man-lin WU, Jie-jun SHI, Tao ZHANG, Yin QIN, Shu-yu LI, Xi-yan HAO, Zhi-gang WANG, Dong-jun LIU
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引用次数: 7
摘要
Rheb作为Ras超家族成员之一,是mTOR信号通路的上游调控因子,调控细胞生长、增殖和分化过程。为了研究内蒙古绒山羊(Capra hircus)细胞中Rheb与mTOR的关系,采用RT-PCR扩增了富含Ras的脑内同源基因(Rheb) cDNA。全长555 bp,包含编码184个氨基酸的完整ORF。HM569224)。全cDNA核苷酸序列与羊、牛和人的同源性分别为99%、98%和93%,氨基酸序列与人的同源性分别为98%、97%和97%。生物信息学分析表明,Rheb具有一个Ras家族结构域、两个酪蛋白激酶II磷酸化位点、两个ATP/ gtp结合位点motif a (P-loop)、一个烯丙基结合位点(CAAX box)。采用半定量RT-PCR进行组织特异性表达分析。Rheb基因在所有被测组织中均有表达,且在脑组织中mRNA积累水平最高,表明Rheb在山羊细胞中发挥了重要作用。
Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus)
As one member of the Ras super family, Rheb is an upstream regulator of mTOR signaling pathway, which regulates the process of cell-growth, proliferation and differentiation. In order to study the relationship between Rheb and mTOR in Inner Mongolian Cashmere goat (Capra hircus) cells, Ras homolog enriched in brain (Rheb) gene cDNA was amplified by RT-PCR. It is 555 bp in length and includes the complete ORF encoding 184 amino acids (GenBank accession no. HM569224). The full cDNA nucleotide sequence has a 99% identity with that of sheep, 98% with cattle and 93% with human while their amino acids sequence shares identity with 98, 97 and 97% of them, correspondingly. The bio informatics analysis showed that Rheb has a Ras family domain, two casein kinase II phosphorylation sites, two ATP/GTP-binding sites motif A (P-loop), a prenyl group binding site (CAAX box). Tissue-specific expression analysis performed by semi-quantitative RT-PCR. The Rheb gene was expressed in all the tested tissues and the highest level of mRNA accumulation was detected in brain, suggesting that Rheb played an important role in goat cells.