The Study On The Expression C-KIT And SCF Gene In Goat Skin And Their Relationship With Coat Color

This study aims to investigate the expression level of c-KIT gene and SCF gene (KIT ligand) ingoat skin with different coat color and its relationship with the number of mature melanocytes, and toscreen SNPs markers associated with coat color related traits on c-KIT gene and SCF gene, with aview to provide a theoretical basis for the practice of molecular breeding for coat color related traitsThe existing of different splicing types in SCF gene mRNA was investigated in this study, thereare soluble type (KC794711.1)，transmembrane type (KC794712.1) and transmembrane-similar type(KC794713.1) three splice variantsof were identified in the skin of goat. The TYR gene was chosen asa marker of mature melanocytes, Real-time quantitative PCR technology was used to detect mRNAexpression level of c-KIT, SCF and TYR gene between skin tissue in goat with black and white coatcolor. The mRNA expression in goat skin tissue with black coat color of TYR gene is8.280times thatof the white; c-KIT gene mRNA expression level was1.255times of the white; soluble SCF mRNAexpression levels is0.267times of the white; transmembrane SCF mRNA expression level is0.874times of the white; transmembrane-similar type SCF mRNA expression level is0.103times the white.The sum of the two soluble SCF and transmembrane-similar type SCF mRNA expression levelsshowed a significant difference between black and white. This may be associated with the formationof different coat color.The c-KIT were located in goat skin tissue using immunohistochemistry. In goat skin tissue,c-KIT mainly in the skin epithelial department, infundibulum, inner and outer root sheath of hairfollicle, no significant differences were observed between skin tissue with black and white coat color,which suggested c-KIT might not be the key factor of the formation between blsck and white coatcolor in this study.Primers were designed by referencing the complete genome sequence of bovine from Ensembldatabase. While four varieties of41goats genome was used as a template of PCR. Sequences of goatc-KIT exon1,2,5,6,7,8,14,15,16,17,18,19,20,21, SCF gene exon1,3,4,6,9and adjacentintron sequences both sides of each exon were got by sequencing.32point mutations were found inc-KIT gene by analysis of sequences comparing, in which g.68332351G>A, g.68346732T>G,g.68355062C>T, g.68359335G>A4polymorphic loci were found as cSNPs, of which the first threepositions were non-synonymous mutations and the rest was synonymous mutations. In this study,g.68346732G was only found in light-colored coat individuals, g.68359156T was detected only inindividuals with dark hair, and g.68332351A existed only in Nanjiang Kuaizhang breed individuals.19SNPs and3InDels with g.17658037insertA, g.17621420insertT, g.17621283insertGA were foundin SCF gene, which include one cSNPs on exon6, the mutation might affect maturation process of soluble SCF.The four cSNPs in c-KIT gene were analyzed and it was found that haplotype ATAC and GTATexisted in the individuals with dark coat color, and GGAC can be detected only in light individual.Analyzed with all SNPs of c-KIT gene,20haplotypes was obtained, with a haplotype diversityof0.892, of which15haplotypes existed only in a single population; Tajima test among Liaoningbreed is significant with a D value of2.414. It showed that balance selection within c-KIT gene mightexist in the population. Analyzed with all22polymorphic loci in SCF genes,25haplotypes of SCFgene was obtained in which differentiation factor is0.905, of which19haplotypes existed only in asingle population, but neither of haplotypes had an obviously linkage with traits.