山羊KAP8.2基因的克隆表达及分析

Analysis of Cloning and Expression for Goat KAP8.2 Gene

目的:研究不同品种山羊和驯鹿高甘氨酸/酪氨酸(HGTKAPs)Ⅰ型蛋白KAP8.2在核苷酸和氨基酸水平的差异,以及KAP8.2mRNA在内蒙古白绒山羊皮肤中的表达。方法:以阿尔巴斯白绒山羊KAP8.2设计特异引物,扩增马头山羊、吐根堡奶山羊、藏山羊和驯鹿KAP8.2基因的编码区,克隆测序并分析。体外制备地高辛标记的KAP8.2cRNA探针,通过原位杂交法在山羊皮肤组织切片上进行定位。结果:发现KAP8.2编码区在核苷酸和氨基酸水平的同源性较高,但也有差异,这可能是毛发性质不同的诸多原因之一。KAP8.2mRNA在成年10月份皮肤、胚胎125d皮肤初级和次级毛囊的皮质层均有强烈的表达。结论:KAP8.2是胚胎和成年山羊皮肤初级和次级毛囊的皮质层的组成成分,不同地区、不同品种山羊及驯鹿的KAP8.2在核苷酸和氨基酸水平同源性较高。

Objective： To study the differences of KAP8 - 2 for different kinds of goats and reindeer, and to detect the expression of KAP8 - 2mRNA on the adult and fetal skins of arbes cashmere goat. Method： According to the conservative sequences of announced arbas cashmere goat KAPS. 2 gene designed a pair of primers of gKAPS. 2, then amplified KAPS. 2 coding region of other kinds of goats and reindeer by PCR, the product was cloned into PMD- 19T vector and sequenced. Made KAP8.2 cRNA probe, In situ hybridization with the adult and fetal skins of arbes cashmere goat. Result： Sequence analysis revealed that nucleotides and encode amino acid sequence were high degree of conservation, maybe the differences was one important reason for hair character. In situ hybridization with the KAPS. 2 cRNA probe on skin revealed a strong positive signal in both primary and secondary hair follicles. Conclusion： The nucleotides and encode amino acid sequence of KAPS. 2 were high degree of conservation for different kinds of goats and reindeer. KAPS. 2mRNA expressed in cortex of both primary and secondary hair follicles.