Cloning and Tissue Expression Analysis of MyoD1 Gene in Luchuan Pig

doi:10.7668/hbnxb.20194657

Abstract

Abstract:

Luchuan pig myoblast determining gene 1(MyoD1)was cloned,bioinformatics analysis was performed,and its expression was analyzed in different tissues of Luchuan pigs,to explore the role of MyoD1 gene in the growth and development of skeletal muscle.Published MyoD1 gene of wild boar sequence on NCBI as the template,designed specific primer,cloned the CDS region of MyoD1,and compared the similarity with wild boar,cattle,sheep,human,mouse and rat gene sequence,constructed system tree for bioinformatics analysis by online software.Then,the relative expression of MyoD1 gene in different tissues of Luchuan pigs by RT-PCR.The CDS of MyoD1 gene in Luchuan pig was 960 bp,encoding 319 amino acids,five base mutations.The similarities of MyoD1 genes between Luchuan pig and wild boar,cattle,sheep,human,mouse and rats were 99.2%,93.0%,92.3%,90.0%,84.3% and 84.0%,respectively.Total number of MyoD1 gene atoms in Luchuan pigs was 4 680,molecular mass of the protein was 33.99 ku,molecular formula was C₁₄₅₇H₂₂₉₆N₄₄₂O₄₇₁S₁₄;the instability coefficient was 63.87,indicating a lack of stability;isoelectric point was 5.63,which belongs to the acidic protein;no transmembrane structure existed,found 35 phosphorylation sites and 1 glycosylation site;protein secondary structure was dominated by an irregular coil,which accounting for 60.69%.The MyoD1 gene in Luchuan pigs had the highest expression in the longest dorsal muscle,which was significantly higher than in other tissues,while kidney had the lowest expression.MyoD1 gene was expressed in all tissues of Luchuan pigs and mainly in the longest dorsal muscle,we speculated that MyoD1 gene might play a crucial role in the muscle growth and development of Luchuan pigs.

Key words: Luchuan pig, MyoD1 gene, Bioinformatics analysis, Gene cloning, Organizational expression

GAO Zeren, PAN Pengcheng, XU Wenwen, CHEN Baojian, LIU Mingjun, GUAN Zhihui, XIE Bingkun, QIN Zhaoxian. Cloning and Tissue Expression Analysis of MyoD1 Gene in Luchuan Pig[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(2): 168-173. doi: 10.7668/hbnxb.20194657.

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Figures/Tables 12

Tab.1 Primer information

基因 Genes	引物序列(5'—3') Primer sequence(5'—3')	片段大小/bp Product length
MyoD1-Fa	ATGGAGCTGCTGTCGCCA	1 006
MyoD1-Ra	TCAGAGCACCTGGTAGATAGGGTT
MyoD1-Fb	GTGCAAACGCAAGACCACTA	128
MyoD1-Rb	CGCTGATTCGGGTTGCTAGA
GAPDH-F	CCCGTTCGACAGATAGCCGTA	83
GAPDH-R	AATCCGTTCACTCCGACCT

Fig.1 The CDS sequence amplified production of the MyoD1 gene

Fig.2 The protein sequence alignment of MyoD1 between Luchuan pig and wild boar

Fig.3 Sequence homology alignment of the MyoD1 genes in different species

Fig.4 Phylogenetic evolutionary tree of the MyoD1 gene

Fig.5 The hydrophilic results of MyoD1 protein

Fig.6 MyoD1 protein signal peptide prediction in Luchuan pigs

Fig.7 Transmembrane structure prediction of MyoD1 protein in Luchuan pigs

Fig.8 Prediction of phosphorylation sites of MyoD1 protein in Luchuan pigs

Fig.9 Prediction of the glycosylation sites of MyoD1 protein in Luchuan pigs

Fig.10 Secondary structure prediction of MyoD1 protein in Luchuan pigs

Fig.11 Tissue expression profiles of the MyoD1 gene Different lowercase letters represent significant differences in different tissue(P<0.05).

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