摘要
为探究黔北麻羊基质金属蛋白酶抑制剂1(TIMP1)基因的表达水平和蛋白结构功能。本研究选取3岁两胎次的单、多羔健康黔北麻羊母羊各3只,采集子宫、输卵管、垂体、下丘脑、卵巢5个组织并提取组织中的总RNA反转录为第一链cDNA,克隆了黔北麻羊TIMP1基因CDS区全长序列,对TIMP1蛋白的理化性质、蛋白质结构等进行预测分析,并进行同源性分析和系统进化树构建;同时采用qRT-PCR检测TIMP1基因mRNA在不同产羔性状的黔北麻羊性腺轴组织中的表达水平。结果发现:T-克隆的黔北麻羊TIMP1基因CDS区为624 bp,编码207个氨基酸,与NCBI上山羊mRNA序列相同;蛋白预测分析结果显示:黔北麻羊TIMP1蛋白的分子式为C1020H1581N283O291S19,相对分子质量为23.07364 ku,等电点为8.62,不稳定系数为64.5;遗传进化树分析得出,与其他9个物种相比较,黔北麻羊TIMP1基因与绵羊和牛的遗传距离最近;qRT-PCR结果显示,垂体、卵巢和输卵管在多羔组中的表达高于单羔组(P<0.01)。本研究揭示了TIMP1基因在单、多羔黔北麻羊5个组织中的表达差异性,对TIMP1基因的T-克隆和蛋白的结构功能预测也丰富了相关研究数据,为进一步探索黔北麻羊产羔性状的分子机制提供参考。
To investigate the expression level and protein structure and function of matrix metalloproteinase inhibitor 1(TIMP1)gene in Qianbei Goats,In this study,three healthy lambs from single lamb group and three lambs from multilamb group were selected as test subjects.The total RNA in uterus,fallopian tube,pituitary,hypothalamus and ovarian tissues was extracted and transcribed into the first cDNA and performed qRT-PCR.The full-length sequence of CDS region of TIMP1 gene was cloned,The results showed that the CDS region of TIMP1 gene was 624bp,encoding 207 amino acids,which was identical with the mRNA sequence of goats landed on NCBI.The relative molecular mass was 23.07364 ku.The isoelectric point is 8.62,and the acid and base properties are alkaline.The instability coefficient is:64.5.The results showing that:the CDS region of TIMP1 gene in Qianbei Goats was 624bp,encoding 207 amino acids,which was identical with the mRNA sequence of goats on NCBI.The results of sequence analysis showing that the molecular formula of TIMP1 protein is C1020H1581N283O291S19.The relative molecular mass is 23.07364 kD.The isoelectric point is 8.62,and the instability coefficient is 64.5.According to the analysis of genetic evolutionary tree,compared with other 9 species,respectively,the genetic distance between TIMP1 gene and sheep and cattle is the closest.qRT-PCR results showing that the multi-lamb group hypophysis,ovary and oviduct were highly expressed in the single lamb group.These results indicate that the differences in the expression of TIMP1 gene in single and multi-lamb Qianbei Goats,and the prediction of the structure and function of TIMP1 protein enriched the relevant research data,providing a reference for further exploring the molecular mechanism of lambing traits in Qianbei Goats.
作者
洪磊
敖叶
周志楠
唐文
阮涌
倪萌萌
陈祥
HONG Lei;AO Ye;ZHOU Zhinan;TANG Wen;RUAN Yong;NI Mengmeng;CHEN Xiang(Key Laboratory of Breeding and Breeding of Plateau Mountain Animals,Ministry of Education,Guizhou University,Guizhou Guiyang 550025,China;College of Animal Science,Guizhou University,Guizhou Guiyang 550025,China)
出处
《中国畜牧杂志》
CAS
北大核心
2020年第7期59-64,共6页
Chinese Journal of Animal Science
基金
国家重点研发计划项目(2018YFD0502005)
国家自然科学基金(31760652)
贵州省优秀青年科技人才项目(黔科合平台人才[2017]5624)。
