哈萨克羊INHβA基因克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2019.05.005

Abstract

Abstract:

This study was aimed to clone the sequence of inhibin beta A (INHβA) in Kazakh sheep,and explore the sequence characteristics of INHβA gene and the structure and function of its encoded protein.One pair of specific primers was disigned based on the sequence of sheep INHβA gene (accession No.:NM_001009458.1) in GenBank,the sequence of INHβA gene in Kazakh sheep was amplified by RT-PCR.INHβA gene was inserted into pMD19-T vector for cloning and sequencing,its nucleotide sequence,amino acids sequence,protein transmembrane,protein modification site and secondary structure,tertiary structure model were predicted and analyzed by bioinformatics methods.The results showed that INHβA gene in Kazakh sheep was 1 278 bp in length and encoded 425 amino acids.The homology of INHβA gene in Kazakh sheep with Ovis aries,Bos taurus,Sus scrofa,Mus musculus,Homo sapiens,Rattus norvegicus,Felis catus,Rytolagus cuniculus and Equus caballus was 98.6%,97.7%,90.4%,87.9%,91.1%,88.2%,91.8%,89.8% and 91.8%,respectively,indicating that INHβA gene was highly conserved among different species.The molecular formula of INHβA protein was C₂₀₇₂H₃₃₂₅N₆₀₃O₆₂₈S₂₆,the molecular weight was 47.57 ku,the theoretical isoelectric point (pI) was 7.87,the instability coefficient was 66.16,the fat solubility index was 78.47,and the hydrophilic value was -0.507.The INHβA protein was an alkaline labile hydrophilic liposoluble protein containing a signal peptide,with no transmembrane structure.The secondary structure prediction showed that alpha helix,beta turn,random coil,and extended chain of INHβA protein accounted for 22.59%,4.47%,54.12% and 18.82%,respectively.The tertiary structure prediction showed that the spatial structure of the protein was mainly alpha helix,and contained β-barrel structures of 8 protein modification sites.This results provided a data reference for the later study of the function of INHβA protein and the effect of INHβA gene on improving the fecundity in Kazakh sheep.

Key words: Kazakh sheep; INHβA gene; cloning; sequence analysis

CLC Number:

XIE Jingang, Baheti Bodai, Kadyken Rizabek, Wurelihazi Hazihan. Cloning and Bioinformatics Analysis of INHβA Gene in Kazakh Sheep[J]. , 2019, 46(5): 1290-1298.

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Cloning and Bioinformatics Analysis of INHβA Gene in Kazakh Sheep

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