膜相关转运蛋白Slc45a2(solute carrier family 45 member 2)调节黑素体中的p H值,进而调节酪氨酸酶活性,在黑色素生成中发挥重要作用。小眼畸形转录因子(microphthalmia-associated transcription factor,MITF)已被证实是Slc45a2的调...膜相关转运蛋白Slc45a2(solute carrier family 45 member 2)调节黑素体中的p H值,进而调节酪氨酸酶活性,在黑色素生成中发挥重要作用。小眼畸形转录因子(microphthalmia-associated transcription factor,MITF)已被证实是Slc45a2的调节因子,然而其调控机制仍待研究。该研究旨在探讨Slc45a2在不同毛色小鼠皮肤是否存在差异表达,以及与毛色形成是否存在相关性。QRTPCR检测显示,Slc45a2在不同毛色小鼠皮肤样品中均有表达,在棕色和灰色小鼠皮肤中Slc45a2的表达量是黑色小鼠皮肤的6.29倍(P<0.01)和1.18倍(P<0.05);Western印迹结果显示,在棕色和灰色小鼠皮肤中Slc45a2蛋白表达量是黑色小鼠皮肤的1.44(P<0.01)和1.03倍;免疫组织化学结果表明,Slc45a2在不同毛色皮肤毛囊的毛基质、内外毛根鞘、毛乳头等区域均有表达。为了进一步了解Slc45a2在黑色素细胞色素沉着的重要作用,将Slc45a2转染到小鼠黑色素细胞并测定黑色素含量,同时检测色素沉着相关基因的表达水平。结果表明,转染Slc45a2与空载组相比黑色素含量明显增加。此外,MITF、TYR、TYRP1和TYRP2蛋白水平分别升高1.31倍、1.37倍、1.63倍和2.21倍,差异显著(P<0.05);TYRP2蛋白显著升高2.47倍(P<0.01)。MITF mRNA显著升高1.64(P<0.05);TYR和TYRP1 mRNA显著升高2.96倍(P<0.01)和8.85倍(P<0.01);TYRP2 mRNA表达量变化不明显。Slc45a2在不同毛色小鼠皮肤中均可有效表达,且差异显著。Slc45a2过量表达,使色素沉着相关基因的表达量及黑色素含量增加。由此表明,Slc45a2通过调控色素的生成,进而影响毛色的形成。展开更多
[Objective]The paper was to study the inheritance of white-partridge feather trait in Wenchang chicken and identify the related genetic variation,so as to provide references for the utilization of white-partridge popu...[Objective]The paper was to study the inheritance of white-partridge feather trait in Wenchang chicken and identify the related genetic variation,so as to provide references for the utilization of white-partridge population of Wenchang chicken and molecular marker-assisted selection of white-partridge feather trait.[Method]The inheritance of white-partridge feather trait was studied by reciprocal cross test.SLC45A2 was selected as a candidate gene based on the genetic characteristics of white-partridge feather and results of previous researches.The variation in the exon domain of SLC45A2 gene was analyzed by re-sequencing method.The relationship between missense mutation/nonsense mutation in the exon domain and white-partridge feather trait was verified.[Result]Reciprocal cross test showed that white-partridge feather trait was associated recessive inheritance compared with yellow-partridge feather.The amplification results indicated that there were five SNPs in SLC45A2 gene,in which three were located in exon domain,including two missense mutations(c.a74g and c.gll54c)and one synonymous mutation(c.c561t).The missense mutation was verified with Wenchang chicken Bawang chicken,Qingyuan partridge chicken,Xinghua chicken and red jungle fowl as materials.The results demonstrated that c.g1154c locus of SLC45A2 gene was completely linked with white-partridge feather trait.[Conclusion]The trait of white-partridge feathers showed recessive inheritance and c.g1154c locus might be the causation.展开更多
基金Supported by Science and Technology Project of Jiangxi Provincial Department of Education(GJJ181080)Start-up Fund of Nanchang Normal University(GJJ181080)"11531"Construction Project of Nanchang Normal University.
文摘[Objective]The paper was to study the inheritance of white-partridge feather trait in Wenchang chicken and identify the related genetic variation,so as to provide references for the utilization of white-partridge population of Wenchang chicken and molecular marker-assisted selection of white-partridge feather trait.[Method]The inheritance of white-partridge feather trait was studied by reciprocal cross test.SLC45A2 was selected as a candidate gene based on the genetic characteristics of white-partridge feather and results of previous researches.The variation in the exon domain of SLC45A2 gene was analyzed by re-sequencing method.The relationship between missense mutation/nonsense mutation in the exon domain and white-partridge feather trait was verified.[Result]Reciprocal cross test showed that white-partridge feather trait was associated recessive inheritance compared with yellow-partridge feather.The amplification results indicated that there were five SNPs in SLC45A2 gene,in which three were located in exon domain,including two missense mutations(c.a74g and c.gll54c)and one synonymous mutation(c.c561t).The missense mutation was verified with Wenchang chicken Bawang chicken,Qingyuan partridge chicken,Xinghua chicken and red jungle fowl as materials.The results demonstrated that c.g1154c locus of SLC45A2 gene was completely linked with white-partridge feather trait.[Conclusion]The trait of white-partridge feathers showed recessive inheritance and c.g1154c locus might be the causation.
