牦牛PRLHR基因的克隆、序列分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2019.12.006

Abstract

Abstract: This study was aimed to clone,analyze sequence and determine the tissue expression of PRLHR gene in yak.The hypothalamus,anterior pituitary,ovary,oviduct and uterus tissues of 5 female yaks and 5 female cattle were collected.The full-length cDNA sequence of PRLHR gene in yak was amplified by RT-PCR techniques,and the characteristics of PRLHR protein in yak was analyzed by bioinformatics methods.The expression of PRLHR gene in yak and cattle were detected by Real-time quantitative PCR.The results showed that the sequence of PRLHR gene in yak was 1 625 bp,which contained 1 113 bp of the CDS region,22 bp of 5'-UTR and 490 bp of 3'-UTR,encoding 370 amino acids.The CDS was high homology with Bos taurus,Bubalus bubalis,Ovis aries,Sus scrofa and Homo sapiens,indicating it was conservative in the evolutionary course.PRLHR protein in yak was an unstable hydrophobic protein with no signal peptide,but had 7 transmembrane domains.There were 13 serine phosphorylation sites,6 threonine phosphorylation sites and 4 tyrosine phosphorylation sites.It was predicted that there were 3 N-glycosylation sites and 10 O-glycosylation sites.The secondary structure of PRLHR protein in yak contained alpha helix (49.19%),random coil (31.89%),extended chain (15.68%) and beta turn (3.24%).The tertiary structure of PRLHR protein in yak had a typical domain of PrRP family in GPCRs superfamily.The results of Real-time quantitative PCR showed that the expression of PRLHR gene in oviduct of yak was significantly higher than that of other tissues (P<0.05).The expression of PRLHR gene in hypothalamus,anterior pituitary,uterus and oviduct tissues of yak was extremely significantly higher than that of cattle (P<0.01).In conclusion,cloning,sequence analysis and tissue expression of PRLHR gene in yak were successfully carried out,which provided a foundation for the further study on the regulatory role of PRLHR gene in yak reproduction.

Key words: yak; cattle; PRLHR gene; cloning; tissue expression

CLC Number:

CHEN Ying, XIA Yi, HE Xiangdong, ZI Xiangdong. Cloning,Sequence Analysis and Tissue Expression of PRLHR Gene in Yak[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(12): 3504-3513.

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Cloning,Sequence Analysis and Tissue Expression of PRLHR Gene in Yak

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