Cloning and Expression Characteristics of Hoxa5 Gene in Goat

doi:10.7668/hbnxb.20192404

Abstract

Abstract: Hoxa5 is a member of the HOX family and plays an important role in the process of growth,development and tumorigenesis.However,there is less research on it in ruminants.In order to obtain the goat Hoxa5 gene sequence,to clarify its expression characteristics and know its expression level in different tissues of goats.The Hoxa5 gene was cloned from the subcutaneous fat tissue of Jianzhou big ear sheep using RT-PCR technology,and its biological characteristics were analyzed by bioinformatics methods.Real-time fluorescent quantitative PCR technology was used to detect the expression of Hoxa5 gene in the heart,liver,spleen,lung,kidney,back,thigh,arm,etc.tissues of Jianzhou big ear sheep.The results showed that the open reading frame of the goat Hoxa5 gene was 813 bp,encoding a total of 270 amino acids.the relative molecular mass of Hoxa5 was 29.283 ku,the isoelectric point was 9.42,and there was no signal peptide and transmembrane domain.This protein was a non-secreted protein;the secondary structure prediction of Hoxa5 protein found that the content of random coils in Hoxa5 protein was the highest,followed by α helix,and β turn accounts for a relatively small number of amino acids;amino acid homology comparison results showed that the Hoxa5 amino acid sequence similarity between Jianzhou big ear sheep and Ovis aries,Bos taurus, Equus caballus, Sus scrofa, Rattus rattus, Cavia porcellus, Mus musculus and Homo sapiens were 100.00%,99.63%,99.26%,99.26%,98.89%,98.89%,98.89% and 98.15%.The results of Real-time fluorescent quantitative PCR showed that Hoxa5 gene was differentially expressed in selected tissues,with the highest expression level in the kidney;Fluorescence localization of pEGFP-Hoxa5 fusion vector transfected into goat subcutaneous adipocyte showed that the gene was mainly located in the nucleus.This study showed that the goat Hoxa5 gene was up-regulated during subcutaneous adipocyte differentiation and the protein was localized in the nucleus. The results suggested that this gene might act as a transcription factor to regulate goat subcutaneous adipocyte differentiation.

Key words: Goat, Hoxa5, Expressing characteristics, Nuclear positioning

CLC Number:

Q78
S826

CHEN Dingshuang, WANG Ruilong, LIN Yaqiu, WANG Yong, ZHU Jiangjiang, LI Xin, ZHANG Hao, LI Yanyan. Cloning and Expression Characteristics of Hoxa5 Gene in Goat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(6): 219-227. doi: 10.7668/hbnxb.20192404.

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