摘要
根据GenBank中报道的绵羊Ghrelin基因mRNA序列设计特异性引物,以甘肃肉用绵羊新品种选育群羔羊皱胃组织RNA为模板,采用RT-PCR技术扩增出绵羊Ghrelin基因并克隆到pMD18-T载体中进行测序验证.用生物信息学软件预测其结构,并构建系统进化树,探讨了绵羊Ghrelin基因的生物学功能.结果表明:克隆的绵羊Ghrelin基因序列共409bp,包含完整的编码区序列,含有1个351bp的完整开放阅读框,编码116个氨基酸;Ghrelin蛋白分子质量为12 975.74u,理论等电点为4.70,信号肽切割点位于第23～24位氨基酸之间,整条多肽链表现为疏水性,是一种典型的跨膜脂溶性蛋白;Ghrelin基因编码产物的二级结构主要以α-螺旋和无规则卷曲为主,主要位于胞外(包括细胞膜),作为一种激素参与机体胁迫应答、免疫应答和转录调控.
In order to amplify the Ghrelin gene of Gansu modern breeding sheep group,specific primers were designed according to GenBank sequence.Total RNA was extracted from the abomasums and cDNA encoding sheep Ghrelin gene was obtained by the reverse-transcription PCR（RT-PCR）.The purified RT-PCR products were cloned into pMD18-T vector and then sequenced.The structure and function of Ghrelin gene-encoded protein was predicted by the bioinformatics software and phylogenetic tree of Ghrelin was constructed.The results showed that the amplified Ghrelin gene was 409 bp,including the complete coding sequence.,which was 351 bp in size,encoded 116 amino acids.The sheep protein of Ghrelin was 12 975.74 u in molecular weight,4.70 in isoelectric point.The protein of Ghrelin had one signal peptide between site 23 and 24,and the whole peptide showed the characterisitic of hydrophobicity,which was a transmembrane protein.The secondary structure of Ghrelin showed that it mainly constituted of α-helix and coils,and it may play an important role in immune response,stress response and transcription regulation in extracellular.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2012年第6期13-19,共7页
Journal of Gansu Agricultural University
基金
农业部现代农业产业技术体系建设专项资金(CARS-39)
甘肃省自然科学基金(3ZS061-A25-077)
甘肃省教育厅高等学校研究生导师科研项目(0702-07)
