【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、San...【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。展开更多
Objective: To find out an effective fixative in immunohistochemistry for high-affinity neurotrophin receptor-tyrosine kinase (Trk) B. Methods: Comparing the results from four groups of adult rats which were fixed by d...Objective: To find out an effective fixative in immunohistochemistry for high-affinity neurotrophin receptor-tyrosine kinase (Trk) B. Methods: Comparing the results from four groups of adult rats which were fixed by different fixatives before the brain sections were processed for TrkB immunohistochemistry. Results: In the four groups, TrkB immunoreactive cells were observed throughout the whole brain, but the intensity of immunoreactive cells and the background staining exhibited a marked difference among the groups. Conclusion: Using 0.3%-0.5% paraformaldehyde in 75% saturated picric acid 0.1 mol/L di-sodium hydrogen phosphate buffer as the fixative may yield the best quality of TrkB immunoreactivity.展开更多
文摘Objective: To find out an effective fixative in immunohistochemistry for high-affinity neurotrophin receptor-tyrosine kinase (Trk) B. Methods: Comparing the results from four groups of adult rats which were fixed by different fixatives before the brain sections were processed for TrkB immunohistochemistry. Results: In the four groups, TrkB immunoreactive cells were observed throughout the whole brain, but the intensity of immunoreactive cells and the background staining exhibited a marked difference among the groups. Conclusion: Using 0.3%-0.5% paraformaldehyde in 75% saturated picric acid 0.1 mol/L di-sodium hydrogen phosphate buffer as the fixative may yield the best quality of TrkB immunoreactivity.
