猪MYNN基因可变剪接体的克隆及表达特性研究

doi:10.11843/j.issn.0366-6964.2018.03.003

Abstract

Abstract:

The aim of this study were to clone alternative splicing isoforms of pig MYNN gene, predict the structures and functions of their coding proteins, and investigate the temporal-spatial expression characteristics of each transcript. The full-length CDS of MYNN was cloned by RT-PCR and the biological characteristics of MYNN protein was analyzed by bioinformatics in Mashen pig. Quantitative real-time PCR was employed to detect the expression patterns of MYNN gene in heart, liver, spleen, lung, kidney, cerebellum, small intestine, stomach, pancreas, longissimus dorsi and fat tissues of Mashen pig, and to study the developmental expression patterns in stomach and longissimus dorsi tissues. Two transcripts of MYNN gene were successfully cloned in present study, and named MYNN-1 (GenBank accession number:KY470829) and MYNN-2 (GenBank accession number:KY670835), respectively. MYNN-1 CDS was composed of 1 830 bp encoding 609 amino acids, which belonged to the stable alkaline soluble protein. Whereas MYNN-2 CDS was composed of 1 746 bp encoding 581 amino acids, which belonged to the unstable alkaline soluble protein. MYNN-2 was 84 bp less than MYNN-1, and lacked the exon6. MYNN-2 was found a C₂H₂ type zinc finger protein domain less than MYNN-1 by the prediction of function domain. The analysis of homology and phylogenetic tree showed that the two transcripts amino acid sequences of pig MYNN gene had high homology and close genetic distance with polar bear, goat, horse, dog, and so on, which proved that the pig MYNN gene was very conservative during evolutionary process. MYNN-1 and MYNN-2 were universally expressed in all pig tissues detected, and there were significant differences in expression among different tissues (P<0.05). The two variants had higher expression levels in stomach, small intestine and pancreas of Mashen pig, and their expression were the lowest in fat tissue. The expression level of MYNN-1 was significantly or extremely significantly higher than that of MYNN-2 in all tissues except kidney (P<0.05, P<0.01), which testified that MYNN-1 was the main variant in pig. The expression of MYNN-1 and MYNN-2 decreased gradually with the increase of age in the stomach of Mashen pig. In longissimus dorsi, the expression of MYNN-1 and MYNN-2 increased firstly and then decreased with the increase of age. In this study, two transcripts of pig MYNN gene were successfully cloned. And it was speculated that MYNN play important role in the process of digestion and absorption, as well as growth and development of skeletal muscle in pig, whereas the specific mechanisms were still remaining to be further elucidated.

CLC Number:

S828.2
Q344

GUO Xiao-hong, LI Meng, GAO Peng-fei, CAO Guo-qing, CHENG Zhi-min, ZHANG Ning-fang, LE Bao-yu, LIU Jian-feng, LIU Xiao-jun, LI Bu-gao. Molecular Cloning on Alternative Splice Variants of Pig MYNN Gene and Their Expression Patterns[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 477-487.

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Molecular Cloning on Alternative Splice Variants of Pig MYNN Gene and Their Expression Patterns

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