畜牧兽医学报2018，Vol.49Issue(3)：477-487,11.DOI:10.11843/j.issn.0366-6964.2018.03.003
猪MYNN基因可变剪接体的克隆及表达特性研究
Molecular Cloning on Alternative Splice Variants of Pig MYNN Gene and Their Expression Patterns
摘要
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 C2 H2 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.关键词
猪/MYNN/可变剪接/生物信息学/表达特性Key words
pig/MYNN/alternative splicing/bioinformatics/expression pattern分类
农业科技引用本文复制引用
郭晓红,李步高,李萌,高鹏飞,曹果清,成志敏,张宁芳,乐宝玉,刘剑锋,刘小军..猪MYNN基因可变剪接体的克隆及表达特性研究[J].畜牧兽医学报,2018,49(3):477-487,11.基金项目
山西省科技创新重点团队项目(201605D131045-24 （）
201705D131028-19) （）
三晋学者项目(2016 （）
2017) （）
