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
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摘要
【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.
【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.
引文
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[6]Wang P Q,Tan Y,Zhang B Y,et al. DNA Polymorphisms of 5'-Flanking Region of Insulin-Like Growth Factor 1 Gene and Their As-sociation with Reproduction Traits in Goats[J]. Journal of IntegrativeAgriculture,2011,10(10):1609-1617.
[7]曹海洋.小尾寒羊ADPN和IGF1基因多态性及其遗传效应分析[D].济南:山东农业大学,2013.
[8]卢振峰.三河牛中GnRHR、FSHR、Leptin、IGF-1基因多态性检测及其与繁殖性状的关联分析[D].银川:宁夏大学,2013.
[9]Burgos S A,Cant J P. IGF-1 stimulates protein synthesis by en-hanced signaling through m TORC1 in bovine mammary epithelialcells[J]. Domestic Animal Endocrinology,2010,38(4):211.
[10]Islam K K,Vinsky M,Crews R E,et al. Association analyses of aSNP in the promoter of IGF1 with fat deposition and carcass merittraits in hybrid,Angus and Charolais beef cattle[J]. Animal Genet-ics,2009,40(5):766-769.
[2]张彩红. IGF-1基因启动子区poly T结构与猪生长性状关联分析[D].广州:华南农业大学,2016.
[3]秦玉梅,李佳玉,杨娴婧,等. IGF-1、OVR基因SNPs位点与玫瑰冠鸡产蛋性能和蛋品质的关联分析[J].中国家禽,2017,39(3):6-11.
[4]Echternkamp S. Insulin-Like Growth Factor-I(Igf-I)Response to O-varian Follicle Ablation in Beef Cattle[EB/OL]. https://www. ars.usda. gov/research/publications/publication/? seqNo115=97256,1999-01-02/2018-01-01.
[5]Kim E S,Shi X,Cobanoglu O,et al. Refined mapping of twinning-rate quantitative trait loci on bovine chromosome 5 and analysis of in-sulin-like growth factor-1 as a positional candidate gene[J]. Journalof Animal Science,2009,87(3):835-843.
[6]Wang P Q,Tan Y,Zhang B Y,et al. DNA Polymorphisms of 5'-Flanking Region of Insulin-Like Growth Factor 1 Gene and Their As-sociation with Reproduction Traits in Goats[J]. Journal of IntegrativeAgriculture,2011,10(10):1609-1617.
[7]曹海洋.小尾寒羊ADPN和IGF1基因多态性及其遗传效应分析[D].济南:山东农业大学,2013.
[8]卢振峰.三河牛中GnRHR、FSHR、Leptin、IGF-1基因多态性检测及其与繁殖性状的关联分析[D].银川:宁夏大学,2013.
[9]Burgos S A,Cant J P. IGF-1 stimulates protein synthesis by en-hanced signaling through m TORC1 in bovine mammary epithelialcells[J]. Domestic Animal Endocrinology,2010,38(4):211.
[10]Islam K K,Vinsky M,Crews R E,et al. Association analyses of aSNP in the promoter of IGF1 with fat deposition and carcass merittraits in hybrid,Angus and Charolais beef cattle[J]. Animal Genet-ics,2009,40(5):766-769.