牦牛RGS1基因克隆及其在发情周期卵巢中的表达

doi:10.11843/j.issn.0366-6964.2017.02.008

摘要/Abstract

摘要：

本研究旨在阐明牦牛RGS1基因的表达特性及其在牦牛发情周期中卵巢组织的表达。通过采集不同发情周期的牦牛卵巢及肌肉、脾、心、肾、胃、小肠、小脑、肝和肺组织，以GenBank上已发布的黄牛RGS1基因序列设计引物，利用RT-PCR方法扩增克隆牦牛RGS1基因及检测其在牦牛各组织中的表达谱，并使用相关生物信息学软件分析RGS1基因的结构和功能。采用实时荧光定量PCR法检测RGS1基因在牦牛发情周期卵巢中mRNA的表达水平。结果表明，本试验克隆得到牦牛RGS1基因718 bp的cDNA序列。同源性与进化树分析表明，牦牛RGS1核苷酸序列与黄牛、野牦牛等大多数哺乳动物的遗传距离较近，其在进化进程中相对较保守。牦牛RGS1基因CDS区为591 bp，编码196个氨基酸残基，相对分子质量为22.48 ku，不含跨膜结构和信号肽，为亲水性蛋白和非分泌蛋白。蛋白二级结构和三级结构以α-螺旋和自由卷曲结构为主。RGS1基因在所选牦牛组织样品中均有表达，其中在小肠、小脑、心和胃中表达量最高。荧光定量PCR结果显示，牦牛黄体期卵巢中RGS1 mRNA的表达量极显著高于卵泡期和红体期（P<0.01），卵泡期中RGS1 mRNA表达水平高于红体期，但差异不显著（P>0.05）。本研究成功克隆了RGS1基因及其在牦牛卵巢3个时期中mRNA的差异性，表明该基因参与发情周期卵巢的内分泌活动调控。

关键词: 牦牛, RGS1, 卵巢, 克隆, 表达

Abstract:

The aim of this study was to clarify the expression characteristics of yak RGS1 gene and its expression pattern in ovaries and other tissues on the estrous cycle. The primer was designed based on the GenBank published Bos taurus RGS1 gene sequences, and the expression pattern of RGS1 was detected by RT-PCR in yak various tissues which include muscle, spleen, heart, kidney, stomach, small intestine, cerebellum, liver, lung and the ovarian during in different periods. The structure and function of RGS1 were analyzed by relate bioinformatics software. The mRNA expression level of RGS1 in different periods of yak estrus cycle were detected by qRT-PCR. The results showed that the yak RGS1 gene contained a 718 bp cDNA fragment, and had high homology with other mammals in nucleotide sequence through sequence alignment and phylogenetic tree, indicating that RGS1 is relatively conservative in the evolutionary process. The length of the yak RGS1 gene was 591 bp in CDS, encoding 196 amino acids, and the molecular weight was of 22.48 ku. The protein was predicted to be hydrophilic, non-secretory, lacking cross-membrane and signal peptide. Its secondary structure and tertiary structure was mainly composed of random α-helix and coil. RGS1 gene was expressed in various tissues of yak, and high abundance in small intestine, cerebellum, heart and stomach. qRT-PCR results showed that the mRNA expression level of RGS1 was significantly higher in luteal phase ovarian of yak than that in follicular phase and red stage（P<0.01). Meanwhile, the mRNA expression level of RGS1 was higher in follicular phase than that in red stage ovarian, but no significant difference (P>0.05). The RGS1 was significant differences expression among the 3 periods of yak ovarian in estrus cycle, which indicated that RGS1 involved in the estrous cycle of ovarian reproductive endocrine activities regulation.

Key words: yak, RGS1, ovary, cloning, expression

中图分类号:

S829.9

胡嘉嘉，熊显荣,石仙，蔡雯祎，兰道亮，李键. 牦牛RGS1基因克隆及其在发情周期卵巢中的表达[J]. 畜牧兽医学报, 2017, 48(2): 252-259.

HU Jia-jia, XIONG Xian-rong, SHI Xian, CAI Wen-yi, LAN Dao-liang, LI Jian. Cloning and Expression of RGS1 Gene in Yak Ovary during the Estrous Cycle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 252-259.

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