光唇鱼MSTN基因克

发布时间：2018-05-16 18:04

本文选题：光唇鱼 + 肌肉生长抑制素基因(MSTN)　；参考：《农业生物技术学报》2017年05期

【摘要】：肌肉生长抑制素(myostatin,MSTN)是骨骼肌生长的负调控因子,MSTN基因在遗传育种上有很大的潜在应用价值。光唇鱼(Acrossocheilus fasciatus)是一种具有较大开发前景的经济鱼类,但其生长速度较慢。为了解光唇鱼MSTN基因的特征并为生长性状标记辅助选择提供基础资料,本研究克隆了光唇鱼MSTN c DNA和基因组DNA序列、检测了组织表达情况并进行了单核苷酸多态性(single nucleotide polymorphism,SNP)位点筛查。结果表明,光唇鱼MSTN c DNA序列全长2 163 bp,含有一个1 128 bp的开放阅读框(open reading frame,ORF)、编码375个氨基酸,预测的编码蛋白具有脊椎动物MSTN的序列特征(如含有1个蛋白酶水解位点RIRR、C端有9个保守的半胱氨酸残基)。基于MSTN氨基酸序列的比对显示不同鱼类之间MSTN序列同源性较高(80%),而鱼类与其他脊椎动物同源性较低(65%左右);光唇鱼与其他鲤科鱼类的MSTN基因高度同源(96.5%)。系统进化树中鱼类聚为一枝、其他脊椎动物聚为另一枝,光唇鱼MSTN位于鲤科鱼类的小簇中。半定量RT-PCR检测显示,MSTN m RNA在光唇鱼脑和肌肉中高表达、肠和肝脏中有微弱表达、其他组织中均未见表达。光唇鱼MSTN基因的DNA序列全长5 828bp,含有3个外显子和2个内含子,在第二内含子中检测到10个SNP位点,但与生长性状(体重、体长)不存在显著相关性(P0.05)。研究结果为探究光唇鱼MSTN基因的功能和筛选生长性状相关辅助育种标记提供了参考资料。
[Abstract]:The MSTN gene, a negative regulatory factor for skeletal muscle growth, has great potential application value in genetics and breeding. Acrossocheilus fasciatus is a kind of economic fish with great development prospect, but its growth rate is slow. In order to understand the characteristics of MSTN gene and provide basic data for marker-assisted selection of growth traits, MSTN c DNA and genomic DNA sequences were cloned. Tissue expression was detected and single nucleotide polymorphisms (SNPs) loci were screened. The results showed that the MSTN c DNA sequence was 2 163 BP in length and contained a 1 128 BP open reading frame ORF, encoding 375 amino acids. The predicted protein has the sequence characteristics of vertebrate MSTN (such as 9 conserved cysteine residues at the C terminal of RIRRN containing a protease hydrolysis site). Alignment based on MSTN amino acid sequence showed that the homology of MSTN sequence was high among different fishes, but the homology between fish and other vertebrates was about 65%, and the MSTN gene homology of Lip fish and other Cyprinidae fishes was 96.5%. The fish in phylogenetic tree are clustered into one branch and the other vertebrates in the other branch. The MSTN is located in the small cluster of Cyprinidae. The results of semi-quantitative RT-PCR showed that the expression of MSTN m RNA was high in brain and muscle, weak in intestine and liver, and not in other tissues. The DNA sequence of MSTN gene was 5 828 BP, containing 3 exons and 2 introns. Ten SNP loci were detected in the second intron, but there was no significant correlation with growth traits (body weight, body length). The results provided a reference for exploring the function of MSTN gene and screening related auxiliary breeding markers for growth traits.
【作者单位】：宁波大学教育部应用海洋生物技术重点实验室;
【基金】：国家星火计划项目(No.2011GA701001) 浙江省海洋与渔业项目(浙海渔计[2012]83号) 浙江省科技厅重大科技专项遗传育种项目(No.2012C12907)
【分类号】：Q78;S917.4

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