多浪羊PROKR2基因克隆、生物信息学及组织差异表达分析

doi:10.16431/j.cnki.1671-7236.2024.04.002

Abstract

Abstract: 【Objective】 This study was aimed to clone prokineticin receptor 2 (PROKR2) gene,and detect the expression of PROKR2 gene in different tissues during the initiation of puberty in Dolang sheep,so as to provide a basis for exploring the role of PROKR2 gene in the initiation of puberty in sheep.【Method】 Using the hypothalamic cDNA in Dolang sheep after first puberty as a template,PROKR2 gene was amplified by PCR, cloned and sequenced.DNAMAN software was used to splice the sequencing results,MegAlign software was used to compare the similarity among different species and construct a phylogenetic tree,and the physicochemical property and structural function of PROKR2 protein in Dolang sheep were predicted by bioinformatics software.Real-time quantitative PCR was used to detect the expression of PROKR2 gene in hypothalamus,hypophysis,ovary,oviduct and uterus of Dolang sheep in prepuberty,puberty and postpuberty.【Result】 The cloned sequence size of PROKR2 gene was 2 641 bp,including 5'-UTR 143 bp,3'-UTR 1 343 bp and CDS region 1 155 bp,encoding 384 amino acids,and the similarity was 99.83% with the predicted mRNA sequence in sheep on GenBank (accession No.:XM_004014342.5).The phylogenetic tree showed that the genetic distance of the PROKR2 gene in Dolang sheep was the closest to Capra hircus,and the farthest to Gallus gallus.Bioinformatics analysis showed that PROKR2 protein was a hydrophobic and stable basic protein with 7 transmembrane structures,which belonged to the transmembrane protein,and there were 31 phosphorylation sites,which mainly played a role in the plasma membrane.There was an interaction relationship between proteins such as GnRH1,PROK1,ANOS1 and PROKR2 protein in Dolang sheep.The results of Real-time quantitative PCR showed that the expression of PROKR2 gene in each stage of hypothalamus was significantly higher than that of other tissues (P＜0.05),and the expression of PROKR2 gene was the highest in the postpuberty stage,while there was no significant change in hypophysis (P＞0.05).The expression of PROKR2 gene in uterus in prepuberty was significantly higher than that in puberty and postpuberty (P＜0.05).The expression of PROKR2 gene in ovary and oviduct tissues were significantly higher than those in the prepuberty and postpuberty stages (P＜0.05).【Conclusion】 In this study,the CDS region sequence of PROKR2 gene was successfully cloned,and PROKR2 protein was a hydrophobic stable basic protein containing 7 transmembrane structures,which was mainly expressed in hypothalamus,and the general trend in the ovary and oviduct were first rising and then decreasing,which provided a reference for further exploring its regulation in the initiation of early puberty in sheep.

Key words: Dolang sheep; PROKR2 gene; cloning; puberty; expression

CLC Number:

S826
Q78

HUANG Qiaoyan, LI Wei, WANG Xinkun, XING Feng. Cloning,Bioinformatics and Tissue Differential Expression Analysis of PROKR2 Gene in Dolang Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1339-1348.

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Cloning,Bioinformatics and Tissue Differential Expression Analysis of PROKR2 Gene in Dolang Sheep

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