绵羊ADAMTS-1基因在不同组织间差异表达及第五外显子多态性分析

Differential Expression of Sheep ADAMTS-1 Gene in Different Tissues and Analysis of Exon 5 Polymorphoism

了探究绵羊ADAMTS-1在不同组织间差异表达及ADAMTS-1第五外显子多态性与蒙古羊母羊(单、双羔)、小尾寒羊多羔(多羔)产羔性状相关性分析,本次试验利用实时荧光定量分析法对不同羊品种组织间差异表达量进行分析,及PCR-SSCP法和直接测序法对ADAMTS-1基因第五外显子多态性进行相关性分析。对三个群体绵羊8个组织进行表达量分析可知,卵巢在所有群体中的表达量显著高于其他组织,而其余组织按照表达水平由高往低依次是肾脏、心脏、肝脏、肺脏、脾脏、垂体、下丘脑。试验发现小尾寒羊多羔卵巢的表达量是蒙古羊双羔母羊的1.54倍、蒙古羊单羔母羊的2.61倍。通过对ADAMTS-1基因第五外显子分析可知,在CDS区出现C1506T、G1509A,且均属于同义突变。3种带型分别命名为AA、AB、AC,由显著性分析可知三种基因型均对产羔数不显著。所有绵羊群体x^2检验表明均处于Hardy-weinberg平衡状态,PIC值属于低度中度多态(0.25<PIC<0.5)。试验所得数据表明ADAMTS-1基因对于蒙古羊和小尾寒羊产羔数有重要的影响,初步认定是蒙古羊和小尾寒羊的多胎主效基因。

In order to reveal differential expression of sheep ADAMTS-1 gene in different tissues and analyze the correlation among its exon 5 polymorphism and Mongolia ewes（single and twin births）, and Small Tail Han ewes（multiple births）, we used real-time quantitative PCR（RQ-PCR） to analyze the differential expression of different sheep breeds, and used PCR-SSCP and direct DNA sequencing technique to correlatively analyze polymorphism of ADAMTS-1 gene exon 5. The analysis of expression quantity of 8 organs in 3 sheep breeds indicated that the expression of ovary in all groups was significantly higher than other organs, and the expression level of the rest organs ranking from high to low was kidney, heart, liver, lung, spleen, pituitary, and hypothalamus. The experiment showed that the ovary expression of ADAMTS-1 gene in Small Tail Han ewes（multiple-births） was1.54 and 2.61 times of that in Mongolia ewes（twin births） and Mongolia ewes（single birth） respectively. Based on the ADAMTS-1 exon 5 gene analysis, we found that C1506 T and G1509 A appeared in the CDS region, which both belonged to the synonymous mutation. Three bands were named as AA, AB and AC, and the significant analysis indicated that these three genotypes were not significant on litter size. The results of x^2 fitness test showed that all sheep populations were in Hardy-weinberg equilibrium state, and PIC value belonged to low and moderate polymorphism（0.25〈PIC〈0.5）. The results of this experiment indicated that ADAMTS-1 gene played an important role on the litter size of Mongolia ewes and Small Tail Han ewes, which was preliminarily considered as the prolificacy major gene of the two.