Mitf-M在羊驼皮肤组织的表达与序列分析及免疫组织化学定位  

Expression,Sequence Analysis and Immunohistochemical Localization of Mitf-M Transcription Factor in Alpaca Skin

作　　者：朱芷葳[1] 贺俊平[2] 于秀菊[2] 程志学[2] 董常生[2] 

机构地区：[1]山西农业大学生命科学学院,山西太谷030801 [2]山西农业大学动物科技学院,山西太谷030801

出　　处：《中国农业科学》2012年第4期794-800,共7页Scientia Agricultura Sinica

基　　金：国家自然基金(30571070);山西农业大学科技创新基金(2011008);山西农业大学博士科研启动基金(XB2010002)

摘　　要：【目的】克隆羊驼皮肤组织表达的Mitf-M CDS序列,比较分析羊驼皮肤组织Mitf-M的特征,进一步定位分析Mitf-M蛋白表达的位置,为深入研究羊驼毛色基因表达机制奠定基础。【方法】采用RT-PCR扩增方法分段扩增Mitf-M CDS区,分别利用Clustal X和BioEdit生物学软件对氨基酸序列进行多重序列比较分析,免疫组织化学技术对Mitf-M蛋白定位分析。【结果】羊驼皮肤组织表达的Mitf-M的CDS区由419个氨基酸组成,具有bHLH-zip转录家族的保守结构域,该基因与牛和犬亲缘关系最近,Mitf-M蛋白主要分布于羊驼皮肤组织毛球基底层细胞之间与毛乳头周围的黑色素细胞中。【结论】与其它物种相比较,羊驼皮肤组织表达的Mitf-M蛋白保守性很强,在进化过程中的变异不大,可见Mitf-M为毛发的色素沉积提供重要的物质基础。【Objective】 Cloning,sequence analysis and tissue location of the Mitf-M CDS domain sequence of alpaca skin could provide a foundation for studying coat color expression mechanism.【Method】The CDS domain sequence of Mitf-M was amplified by RT-PCR,multiple sequence alignment of the amino acid sequence was analyzed by Clustal X and BioEdit software.The distribution of Mitf-M protein in alpaca skin was studied by immunohistochemistry.【Result】Mitf-M CDS domain of alpaca is composed by 419 amino acids,with bHLH-zip conserved domain of transcription factor family.Phylogenetic analysis showed that Mitf-M had the highest homology with cattle（Bos Taurus） and dogs（Canis familiaris）.The Mitf-M protein was mainly located in melancytes during the basal cells of hair bulb and around dermis papilla.【Conclusion】 Compared with other species,Mitf-M in alpaca skin are highly conservative,indicating that Mitf-M plays an important role in the transcriptional regulation and pigmentation.

关 键 词：羊驼 皮肤 毛囊 Mitf-M PCR 免疫组织化学 

分 类 号：S829.9[农业科学—畜牧学]