牦牛性激素结合蛋白基因克隆及组织表达研究

doi:10.16431/j.cnki.1671-7236.2019.09.007

Abstract

Abstract: The aim of this study was to investigate the potential role of sex hormone-binding globulin (SHBG) in regulating yak reproduction by clarifying its CDS sequence and expression characteristics in the reproductive axis of female yak (Bos grunniens).Tissue samples of hypothalamus,anterior pituitary,oviduct,ovary and uterus were collected from five adult and health female yaks and cattle,respectively.The SHBG gene-specific amplification primers of yak were designed according to bovine SHBG gene in NCBI.Cloning,bioinformatics analysis and tissue expression of yak SHBG gene were performed by RT-PCR,bioinformatics methods and Real-time PCR,respectively.The result showed that the coding region (CDS) of yak SHBG was 1 206 bp,encoding a 401 amino acids protein which contained a high proportion of leucine (16.5%),glycine (9.7%),proline (8.7%) and serine (9.0%).The formula of SHBG protein was C₁₉₄₄H₃₀₆₁N₅₃₅O₅₆₆S₁₀ and its molecular weight and theoretical isoelectric point were 43.30 ku and 5.63,respectively.Yak SHBG had the highest homology with cattle in nucleotide sequence (99.0%) and amino acid sequence (97.5%).It was a hydrophilicity instability protein contained a signal peptide and a transmembrane region.Random coil and extend chain were mainly in the secondary structure of SHBG,which was consistent with the result of the tertiary structure analysis.The phylogenetic tree showed the closest genetic relationship between yak and cattle.The expression of SHBG gene in yak oviduct was significantly higher than that in other tissues (P<0.05),and the expression among other tissues was not significantly different (P>0.05).The expression of SHBG gene in cattle oviduct was extremely significantly higher than that in yak (P<0.01),and the expression in cattle ovary was significantly higher than that in yak (P<0.05).The expression of SHBG gene in other tissues was not significantly different between yaks and cattle (P>0.05).The research on the SHBG gene sequence and its expression in the reproductive axis of the yak provided a theoretical basis for further studying on the regulation of SHBG on the reproduction in yak.

Key words: yak; SHBG gene; cloning; bioinformatics analysis; tissue expression

CLC Number:

S823.8⁺5

HE Xiangdong, WANG Qin, XIA Yi, CHEN Ying, JIGE Moti, ZI Xiangdong. Cloning and Tissue Expression of Sex Hormone-binding Globulin Gene of Yaks[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(9): 2551-2559.

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Cloning and Tissue Expression of Sex Hormone-binding Globulin Gene of Yaks

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