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1224 bp,相对分子质量为46170.24 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。
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 1224 bp and its relative molecular weight was 46170.24 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.
作者
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
SUN Ruiping;WANG Feng;CHAO Zhe;LIU Hailong;XING Manping;LIU Quanwei;HUANG Lili;ZHENG Xinli;WEI Limin(Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Science, Haikou 571100, China;Hainan Key Lab of Tropical Animal Reproduction & Breeding and Epidemic Disease Research, Haikou 571100, China)
出处
《浙江农业学报》
CSCD
北大核心
2020年第6期978-985,共8页
Acta Agriculturae Zhejiangensis
基金
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)。
