The Asscociation Analysis Of FGF5Gene With Wool Traits In Sheep And Effect Of FGF5on Proliferation Of Sheep Fibroblast

Length is one of key qualities to determine the wool value, so length improvement is animportant goal in sheep breeding. Fibroblast growth factor5(Fibroblast Growth Factor-5,FGF5) is a candidate gene to regulate sheep wool length. In this study, nature wool length andstretched length of278Chinese Merino sheep at Bazhou breeding farm were measured. DNAwas extracted from sheep blood,PCR was conducted to sequence FGF5gene, to find thegene’s variation, and to determine the influence of FGF5variation on sheep wool traits. Onthis basis, a high-throughput HRM FGF5gene genotype analysis techniques system wasconducted to large-scaly detect FGF5mutation, which laid a technical foundation formolecular breeding to improve wool length. Overexpression of FGF5and expression of FGF5,FGF5S, were induced in fibroblasts to analyse FGF5’s function in fibroblasts.The main resultsobtained are as follows:1. By sequencing271Chinese Merino samples, three SNP sites were found. Phenotypesand the three SNPs were analysed in271sheep. We Found Locus1, AA genotype frequencyis0.096, AG type is0.399, GG type is0.505; difference between AA and GA genotype wassignificantly (p <0.01), AA and GG genotype were significantly different (p <0.05), theaverage performance of the three genotypes of wool natural length is: AA <GG <GA. Locus2,AA genotype frequency is0.118, AG type is0.410, GG type is0.472; GG-type natural lengthof the longest wool. Locus3, GG genotype frequency was0.122, TG type is0.417, TT type is0.461; GG type of wool is the longest natural length.2. By adding2.2uM Mg2+and20%of the reference DNA to optimize HRM reactionconditions, which can be clearly distinguished FGF5gene homozygous CC, TT wild type, CTheterozygous. Further analysis by PCR sequencing was conducted to verify the accuracy ofthe HRM in sheep FGF5gene analysis. Initially established HRM technology system to detectsheep FGF5gene SNP provides technical support for the follow-up tests. 3.Cell growth curve showed that overexpression FGF5s can significantly improve SheepFibroblasts in the first24h-72h curve tended to increase; while FGF5result showed its growthcurve in the first24-72h less than FGF5s, FGF5s competitive binding the receptor of FGF5,inhibiting function of FGF5.In summary, the study found genetic variation of Chinese Merino in FGF5gene andfound that the promoter region of the specific SNP and there is a significant correlationbetween the SNP and wool length. Establishment of a suitable FGF5gene mutation analysisin HRM technology system provides technical base for large-scale analysis of FGF5genemutation detection. At the cellular level, initially proved FGF5gene can significantly affectthe growth of fibroblasts.