屯昌猪PDK4基因克隆及其组织表达分析

doi:10.3969/j.issn.1004-1524.2020.06.05

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (6): 978-985.DOI: 10.3969/j.issn.1004-1524.2020.06.05

屯昌猪PDK4基因克隆及其组织表达分析

孙瑞萍^1,2, 王峰¹, 晁哲^1,2, 刘海隆^1,2, 邢漫萍¹, 刘圈炜^1,2, 黄丽丽¹, 郑心力¹, 魏立民^1,2,*

1.海南省农业科学院畜牧兽医研究所,海南海口 571100;
2.海南省热带动物繁育与疫病控制重点实验室,海南海口 571100

收稿日期:2019-12-12 出版日期:2020-06-25 发布日期:2020-06-24
通讯作者: *魏立民,E-mail:liminedu@126.com
作者简介:孙瑞萍(1982—),女,山西运城人,博士,副研究员,主要研究方向为家畜遗传育种与繁殖。E-mail:ruiping937@126.com
基金资助:
国家自然科学基金(31560628,31760646); 海南省省属科研院所技术开发研究专项(KYYS-2018-06); 中央引导地方科技发展专项资金(ZY2019HN01)

Cloning and tissue expression analysis of PDK4 gene in Tunchang pig

SUN Ruiping^1,2, WANG Feng¹, CHAO Zhe^1,2, LIU Hailong^1,2, XING Manping¹, LIU Quanwei^1,2, HUANG Lili¹, ZHENG Xinli¹, WEI Limin^1,2,*

1. Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Science, Haikou 571100, China;
2. Hainan Key Lab of Tropical Animal Reproduction & Breeding and Epidemic Disease Research, Haikou 571100, China

Received:2019-12-12 Online:2020-06-25 Published:2020-06-24

摘要/Abstract

摘要： 为获得屯昌猪丙酮酸脱氢酶激酶4(pyruvate dehydrogenase kinase 4,PDK4)基因的编码序列(coding sequence,CDS)并分析其分子结构特征,研究屯昌猪、杜洛克猪及其杂交F1代不同组织中的PDK4基因表达水平,本研究以Genbank上公布的猪PDK4基因序列(登录号:NM_001159306)为参考设计引物,通过RT-PCR扩增、测序、拼接获得屯昌猪PDK4基因CDS区。结果显示:屯昌猪PDK4基因CDS区长度1 224 bp,相对分子质量为46 170.24 u,既没有跨膜结构也不存在信号肽,属于非分泌型蛋白,主要在线粒体中发挥作用,预测存在2个潜在的糖基化位点和33个磷酸化位点,α螺旋区域在预测的二级结构中占比最大。实时荧光定量PCR检测结果显示,PDK4基因在屯昌猪背最长肌中表达量最高,且极显著高于杜洛克猪及其杂交F1代猪。

关键词: 屯昌猪, PDK4基因, 克隆, 组织表达, 杜洛克猪, 实时荧光定量PCR

Abstract: To obtain the coding sequence (CDS) of PDK4 gene in Tunchang pigs, analyze its molecular structure characteristics, study the expression levels of PDK4 mRNA in different tissues of Tunchang pigs, Duroc pigs and their hybrid F1 generation, pig PDK4 gene (Genbank No.: NM_001159306) was used as the reference primer, and the PDK4 CDS region of Tunchang pigs was obtained by RT-PCR amplification, sequencing and splicing. The results showed that the coding region of PDK4 gene was 1 224 bp and its relative molecular weight was 46 170.24 u. There was neither transmembrane structure nor signal peptide in PDK4 protein, and it was a non-secretory protein which mainly played a role in mitochondria. It was predicted that there could be two potential glycation sites and thirty-three phosphorylation sites, and the helical region occupied the largest proportion in the predicted secondary structure. The results of fluorescence quantitative PCR showed that in different tissues of Tunchang pig, the expression level of PDK4 gene was the highest in the longest dorsal muscle. The expression level of PDK4 gene in Tunchang pig's longest dorsal muscle was significantly higher than Duroc pig and its hybrid F1 generation.

Key words: Tunchang pig, PDK gene, cloning, tissue expression, Duroc pigs, real-time fluorescent quantitative PCR

中图分类号:

S828

孙瑞萍, 王峰, 晁哲, 刘海隆, 邢漫萍, 刘圈炜, 黄丽丽, 郑心力, 魏立民. 屯昌猪PDK4基因克隆及其组织表达分析[J]. 浙江农业学报, 2020, 32(6): 978-985.

SUN Ruiping, WANG Feng, CHAO Zhe, LIU Hailong, XING Manping, LIU Quanwei, HUANG Lili, ZHENG Xinli, WEI Limin. Cloning and tissue expression analysis of PDK4 gene in Tunchang pig[J]. , 2020, 32(6): 978-985.

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屯昌猪PDK4基因克隆及其组织表达分析

Cloning and tissue expression analysis of PDK4 gene in Tunchang pig

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