牦牛PLIN2基因的克隆及其在卵巢不同发情阶段的表达

doi:10.16431/j.cnki.1671-7236.2017.03.023

摘要/Abstract

摘要：

试验旨在研究牦牛PLIN2基因的序列特征和表达特性，并分析其表达与不同发情时期的卵巢的相关性。根据GenBank中已知的普通牛序列，设计克隆引物，经RT-PCR扩增得到牦牛PLIN2基因序列。RT-PCR检测PLIN2基因在各个组织中的表达量，实时荧光定量PCR分析卵巢不同发情时期的表达量。结果表明，牦牛PLIN2基因的编码区长为1 353 bp，共编码450个氨基酸，为无跨膜结构的非分泌蛋白。与GenBank中已知的普通牛序列同源性达到99.4%，表明PLIN2基因在进化过程中相当保守。在牦牛的各个组织中PLIN2分布广泛，但不同组织中的表达有差异，卵巢、乳腺、胃和肾脏中相对较多，而肺脏中几乎没有表达。实时荧光定量PCR结果显示，在卵巢的不同发情时期PLIN2都有表达，且在黄体期的表达量最高，表明PLIN2基因在牦牛发情周期中起着不可忽视的作用。

关键词: 牦牛; PLIN2基因; 克隆; 组织表达; 实时荧光定量PCR

Abstract:

The purpose of this study was to research the sequence characteristics and the expression of PLIN2 gene, and to analyze the relationship between the expression and the ovaries in different estrus periods. According to the gene sequence of bovine in GenBank, the primer was designed. The PLIN2 gene of yak was cloned by RT-PCR, and the expression quantity of the PLIN2 gene in different tissues of yak was detected by the semiquantitative RT-PCR. Meanwhile,the differential expression of the PLIN2 gene in different ovarian estrus periods of yak was detected by quantitative Real-time PCR. The results showed that the length of the PLIN2 gene was 1 353 bp in CDS, encoding 450 amino acids. The PLIN2 protein was predicted to be non-seretory and lacking cross-membrane domain. It was 99.4% homology identity to that of corresponding cDNA from bovine, which showed that the PLIN2 gene was quite conservative in the process of evolution. The expession profile of PLIN2 was wide in yak tissues, but there were some differences in different tissues. The expression quality was higher in kidney, breast and ovary, while it was almost nothing in lung. The result of quantitative Real-time PCR indicated that yak PLIN2 gene was expressed in different ovarian estrus periods, and the highest expression during the luteal phase, which indicated the PLIN2 gene played an essential role in yak estrous cycles.

Key words: yak; PLIN2 gene; clone; expression in tissue; quantitative Real-time PCR

中图分类号:

Q785

蔡雯祎, 陈通, 石仙, 陈伟明, 胡嘉嘉, 熊显荣, 兰道亮, 李键. 牦牛PLIN2基因的克隆及其在卵巢不同发情阶段的表达[J]. 《中国畜牧兽医》, 2017, 44(3): 782-789.

CAI Wen-yi, CHEN Tong, SHI Xian, CHEN Wei-ming, HU Jia-jia, XIONG Xian-rong, LAN Dao-liang, LI Jian. Cloning of PLIN2 Gene of Yak and Its Expression in Ovary during Different Estrus Periods[J]. , 2017, 44(3): 782-789.

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