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天祝白牦牛 IGF-1基因克隆、生物信息学及差异表达分析
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引用本文:常永芳,包鹏甲,张永峰,付东海,吴晓云,褚 敏,阎 萍.天祝白牦牛 IGF-1基因克隆、生物信息学及差异表达分析[J].西北农业学报,2019,28(12):1934~1941
DOI:10.7606/j.issn.1004-1389.2019.12.004
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作者单位
常永芳,包鹏甲,张永峰,付东海,吴晓云,褚 敏,阎 萍 (1.西北民族大学 生命科学与工程学院兰州 7300302.中国农业科学院 兰州畜牧与兽药研究所甘肃省牦牛繁育工程重点实验室兰州 730050) 
基金项目:国家现代农业(肉牛牦牛)产业技术体系建设专项(CARS-37);中国农业科学院创新工程(CAASASTRP-2014-LIHPS-01)。
中文摘要:旨在获得天祝白牦牛 IGF-1基因序列,分析其分子结构特征及在牦牛毛囊发育周期中的表达规律,为解析白牦牛毛囊发育及被毛生长提供理论依据。以天祝白牦牛体侧皮肤为试验材料,通过PCR扩增及测序技术克隆 IGF-1基因CDS区序列,并与黄牛序列比对,进行生物信息学分析;采用实时荧光定量PCR(qPCR)技术检测天祝白牦牛毛囊发育不同时期 IGF-1基因的mRNA表达水平。结果表明,天祝白牦牛 IGF-1基因的CDS区长465 bp,编码154 个氨基酸; IGF-1基因编码蛋白的分子质量为17 065. 81 ku,理论等电点为9.36;蛋白质二级结构以无规卷曲(57.79%)和α-螺旋(27.27%)为主,序列分析表明,天祝白牦牛IGF-1蛋白为不稳定亲水蛋白;qPCR检测结果表明, IGF-1基因在牦牛毛囊发育的生长期、退行期和休止期均有表达,生长期和退行期表达量显著高于休止期。
中文关键词: IGF-1基因  天祝白牦牛  克隆  生物信息学
 
Cloning, Bioinformation Analyzing, and Expressional Detecting for IGF-1 in Tianzhu White Yak
Abstract:The purpose of this study is to obtain the sequence and molecular structure of IGF-1 gene from Tianzhu white yak, to analyze expression pattern in the development cycle in yak hair follicles, to provide a theoretical basis for analyzing the development of hair follicles and hair growth of Tianzhu white yak. Therefore, in the experiments, the skin of Tianzhu white yak was chosen as experimental material, and the full-length coding sequence(CDS) of yak IGF-1 gene was cloned. The cloned IGF-1 gene CDS was compared with the cattle sequence and analyzed by related softwares and IGF-1 mRNA expression of Tianzhu white yak hair follicle development was detected in the anagen, catagen and telogen by real-time quantitative PCR (qPCR). The results showed that the CDS of IGF-1 was 465 bp in lengeth, encoded 154 amino acid with the molecular mass of protein of 17 065.81 ku and the theoretical isoelectric point was 9.36. The secondary structure of IGF-1 protein mainly contained random coil (57.79%) and α-helix (27.27%). The sequence analysis showed that IGF-1 protein of Tianzhu white yak was an unstable hydrophilic protein. The results of qPCR showed that IGF-1 gene was expressed in the anagen, catagen and telogen of yak hair follicles, and the expression was significantly higher in the telogen thanthose of anagen and catagen . This study revealed the molecular structure characteristics of the IGF-1 encoding sequence of Tianzhu white yak and the expressional characteristics in the development cycle of yak hair follicle, providing theoretical basis for the biological function that IGF-1 regulates the hair follicle development in Tianzhu white yak.
keywords: IGF-1 genes  Tianzhu white yak  Clone  Bioinformatics
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