林麝繁育性状候选基因IGF-1的生物信息学分析及遗传效应研究

Bioinformatics Analysis and Genetic Effect Research of Candidate Gene IGF-1 for Breeding Traits on Forest Musk Deer

【目的】本研究建立了林麝标记辅助选择技术体系,为林麝的品系选育提供理论依据。【方法】通过基因克隆研究林麝胰岛素样生长因子(IGF-1)的基本功能和蛋白质结构,采用PCR-SSCP技术检测IGF-1基因在林麝中的多态性,分析其与产仔数的关联效应。【结果】①IGF-1基因序列全长465 bp,编码153个氨基酸残基,经生物信息学分析,发现该蛋白含有15个磷酸化位点,是一个分泌性蛋白,大部分位于细胞膜上,含有Insulin家族典型的保守结构功能域;②本试验设计IGF-1基因的PCR-SSCP扩增片段长度均为300 bp左右,引物扩增片段用10%的聚丙烯酰胺凝胶进行检测,发现扩增片段没有特异性条带。为证明PCR-SSCP扩增结果,选择其中一条引物对所得的毛发样品进行扩增,测序发现所有PCR产物的序列完全一致,没有多态性。【结论】研究表明不能选择IGF-1基因作为林麝的分子遗传标记,但为林麝产香性能的分子遗传标记选育提供了基本思路。

【Objective】This study established the technical system of musk deer marker assisted selection to provide a theoretical basis for the breeding of forest musk deer.【Method】The basic functions and protein structure of forest musk deer insulin-like growth factor(IGF-1)by gene cloning were analyzed,the polymorphism of IGF-1 gene by PCR-SSCP technology was detected,and its correlation effect on the number of litter was explored.【Result】(i)The sequence of IGF-1 gene was 465 bp,encoding 153 amino acid residues,and the bioinformatics analysis revealed that the protein contains 15 phosphorylation sites,a secreted protein mostly located in the cell membrane,containing a conserved Insulin family of typical domain;(ii)The length of PCR-SSCP amplified fragment of IGF-1 gene was 300 bp,the primer amplified fragment was detected by 10%polyacrylamide gel,and it was found that there was no specific band in the amplified fragment.In order to prove the result of PCR-SSCP amplification,one of the primers was selected to amplify the hair samples,and sequencing showed that all PCR products had a complete sequence and no polymorphism.【Conclusion】This research showed that the IGF-1 gene was not chosen as a molecular genetic marker of forest musk deer,which would provide a basic idea for the breeding of molecular genetic markers of incense production performance of forest musk deer.