摘要
[目的]探究酪氨酸酶相关蛋白1b(tyrosinase-related protein 1b,TYRP1b)基因变异与斑马鱼体色性状的相关性.[方法]以体色表型存在显著差异的红色、黄色和蓝色斑马鱼共计219个样本的肌肉组织DNA为模板,设计3对引物,采用PCR扩增、Sanger测序技术筛查TYRP1b基因单核苷酸多态性(single nucleotide polymorphisms,SNPs),并将SNPs与斑马鱼体色性状进行关联分析.[结果]获得的斑马鱼TYRP1b基因外显子2、部分内含子3、外显子4、内含子4、外显子5、内含子5、外显子6和外显子7长度分别为134、212、168、135、180、113、150、171 bp.SNPs筛查显示,3种体色斑马鱼群体共筛查到18个SNPs,部分内含子3存在11个SNPs位点:g.2125G＞A、g.2150G＞A、g.2152G＞T、g.2161C＞A、g.2175A＞T、g.2180G＞T、g.2185A＞T、g.2192A＞C、g.2200A＞T、g.2201A＞C 和 g.2213T＞C;内含子 4 检测到 4 个 SNPs 位点:g.7C＞A、g.65A＞G、g.84T＞A 和g.103C＞T;内含子5存在1个SNPs位点:g.61G＞T;外显子4、6分别存在1个SNPs位点:g.125G＞T和g.80T＞A,且均为同义突变;外显子2、5和7均未检测到SNPs.关联分析表明,TYRP1b基因内含子3中g.2152G＞T、g.2175A＞T、g.2180T＞G、g.2192A＞C、g.2200A＞T、g.2201A＞C位点、内含子 4 中 g.65A＞G位点及外显子6中g.80T＞A位点的基因型均与斑马鱼体色性状极显著相关(P＜0.01).[结论]斑马鱼TYRP1b基因内含子3、4及外显子6存在8个与体色表型存在关联的SNPs,可能影响斑马鱼体色性状或与之紧密连锁.
Abstract
[Objective]Correlation between the mutation sites of tyrosinase related protein lb gene,TYRP1b,and body color traits of Danio rerio was investigated.[Method]DNA of muscles from zebrafish of three significantly different body color phenotypes were used as the templates.Single nucleotide polymorphisms(SNPs)of the genes were tested by PCR and Sanger direct sequencing to determine the relationship between the mutation sites of TYRPib and traits of zebrafish body color.[Result]The exon 2,partial intron 3,exon 4,intron 4,exon 5,intron 5,exon 6,and exon 7 of TYRP1b were 134 bp,212 bp,168 bp,135 bp,180 bp,113 bp,150 bp,and 171bp in length,respectively.Eighteen SNPs were found in 219 zebrafish with body color of red,yellow,and blue.Of them,11(i.e.,g.2125G＞A,g.2150G＞A,g.2152G＞T,g.2161C＞A,g.2175A＞T,g.2180G＞T,g.2185A＞T,g.2192A＞C,g.2200A＞T,g.2201A＞C,and g.2213T＞C)were located in partial intron 3,5(i.e.,g.7C＞A,g.65A＞G,g.84T＞A,and g.103C＞T)in intron 4 and g.61G＞T in intron 5,and the synonymous mutation g.125G＞T in exon 4 and g.80T＞A in exon 6.No SNPs were detected in exons 2,5,and 7.An association analysis showed that the genotypes of g.2152G＞T,g.2175A＞T,g.2180T＞G,g.2192A＞C,g.2200A＞T,and g.2201A＞C in intron 3,g.65A＞G in intron 4,and g.80T＞A in exon 6 of TYRP1b significantly correlated with zebrafish body color(P＜0.01).[Conclusion]It appeared that 8 SNPs in introns 3,4,and exon 6 of TYRP1b in D.rerio were associated with body color phenotype of the fish.
基金项目
国家自然科学基金项目(32060274)
贵州省科技计划项目(黔科合基础-ZK[2022]一般559)
贵州省科技支撑项目(黔科合支撑[2023]一般081号)
贵州省高层次创新型"千"层次人才培养项目(2024-[2022]-048)
贵州省重点实验室项目(黔科合平台人才[2020]2003号)
维普
万方数据