Cloning and Expression Analysis of PHKG1 Gene in Goat

doi:10.7668/hbnxb.20190880

Abstract

Abstract: The aim of this experiment was to obtain the sequence of goat PHKG1,to clarify its biological characteristics,and to elucidate the tissue expression level and cell expression profiles of subcutaneous and intramuscular adipocytes. Jianzhou Big-eared goats were selected as experimental materials,the heart,liver,spleen,kidney,lung,longissimus muscle,subcutaneous fat were harvested after slaughtering,and the total RNA was extracted from these tissues. The sequence of PHKG1 gene was cloned by Real-time PCR(RT-PCR),the biological characteristics were analyzed by online tools,and the tissue and cell temporal expression of PHKG1 was detected by Real-time quantitative PCR (qPCR). The results showed that the cloned PHKG1 gene sequence was 1 233 bp in length,of which the CDS region was 1 164 bp. The PHKG1 gene encoded 387 amino acids,forming an unstable hydrophilic acidic nontransmembrane protein with no signal peptide,and subcellular localization indicated that it was mainly present in the cytoplasm. The amino acid sequence similarity between goats and sheep,cattle,pigs,horses and humans was above 90%,indicating that PHKG1 was highly conserved in different species. Constructing phylogenetic tree showed that goats and sheep were in the same branch,in line with the evolutionary laws of species. The PHKG1 gene was widely expressed in goat-tissues and was the highest in the longissimus muscle(P <0.01)and the expression levels in intramuscular adipocytes at 60 h and subcutaneous adipocytes at 96 h of adipogenic differentiation were significantly(P<0.05)and extremely significantly(P<0.01)higher than those in the preadipocytes. This study laid the foundation for further study of the role of PHKG1 in the differentiation of goat adipocytes.

Key words: Goat, PHKG1, Gene cloning, Tissues expression, Temporal expression

CLC Number:

Q78
S826

LI Xin, HE Xiaofang, ZHANG Hao, ZHENG Jianying, WANG Yuxue, LIN Yaqiu, WANG Yong, ZHU Jiangjiang. Cloning and Expression Analysis of PHKG1 Gene in Goat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 195-202. doi: 10.7668/hbnxb.20190880.

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