新西兰白兔MSTN基因克隆、生物信息学分析及表达谱构建

doi:10.16431/j.cnki.1671-7236.2021.10.001

Abstract

Abstract: Myostatin (MSTN) was a master regulator of skeletal muscle development. In order to obtain the sequence and expression pattern of MSTN gene in New Zealand White rabbits, the MSTN gene sequence was amplified and cloned from leg muscle tissue of New Zealand White rabbit by RT-PCR and then was analyzed by online bioinformatics software. The expression level of MSTN gene in different tissues and different periods of leg muscle was detected by Real-time quantitative PCR. The results showed that the CDS region of MSTN gene in New Zealand White rabbit was 1 128 bp in length, encoding 375 amino acids, and had 95.9%, 94.9%, 94.9%, 91.7%, 91.5%, 91.3% and 84.2% similarity with the nucleotide sequences of Homo sapiens, Sus scrofa, Equus caballus, Mus musculus, Canis lupus familiaris, Bos taurus, Gallus gallus published in GenBank, respectively. MSTN protein was an acidic and hydrophilic secretory protein, without transmembrane structure and contains a signal peptide. It mainly distributed in endoplasmic reticulum and mitochondria. The prediction results of secondary structure and tertiary structure of MSTN protein were consistent, which were mainly composed of random coil, followed by alpha helix and extended chain. It was a mixed protein. Protein interaction prediction found that MSTN protein interacted with PAX7, MYOG, IGF1, FST, Smad3 and Smad2 proteins related to muscle growth and development. Results of expression analysis in different tissues showed that MSTN gene was widely expressed in heart, liver, spleen, lung, kidney, leg muscle, uterus and stomach of New Zealand White rabbits. The expression level in leg muscle was the highest, which was extremely significantly higher than that in other tissues (P<0.01), followed by kidney, and the lowest expression in liver. Results of expression analysis in leg muscle tissue at different stages showed that the expression of MSTN gene in 180-day-old leg muscle tissue of New Zealand White rabbits was significantly higher than that in 90- and 240-day-old (P<0.05). The results laid a foundation for further study on the regulatory mechanism of MSTN gene on muscle growth and development in rabbits.

Key words: New Zealand White rabbits; MSTN gene; bioinformatics analysis; Real-time quantitative PCR; expression

CLC Number:

S829.1

JIANG Yixuan, WANG Yaling, XING Shanshan, SONG Guohua, XU Huifen, LI Ming. Cloning, Bioinformatics Analysis and Expression Profile Construction of MSTN Gene in New Zealand White Rabbits[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3501-3511.

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Cloning, Bioinformatics Analysis and Expression Profile Construction of MSTN Gene in New Zealand White Rabbits

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