山羊KAP6-1.2基因的克隆表达及分析  

Analysis of Cloning and Expression for Goat KAP6-1.2 Gene

作　　者：张俊霞[1] 王利[2] 尹俊[1] 李金泉[2] 

机构地区：[1]内蒙古农业大学生物工程学院,内蒙古呼和浩特010018 [2]内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特010018

出　　处：《生物技术》2007年第3期20-23,共4页Biotechnology

基　　金：国家自然科学基金地区联合资助项目(30660122);内蒙古自然科学基金项目(200308020407);内蒙古自然科学基金重大项目(200408020401)

摘　　要：目的:研究角蛋白关联蛋白KAP6-1.2对不同品种羊绒和羊毛在核苷酸和氨基酸水平上的影响及在山羊皮肤中的定位表达。方法:以阿尔巴斯白绒山羊KAP6-1.2 cDNA设计特异引物,扩增5种山羊的KAP6-1.2基因的编码区,克隆并测序。制备KAP6-1.2 cRNA探针,通过组织切片原位杂交在皮肤中进行定位表达分析。结果:6种山羊的KAP6-1.2在核苷酸和氨基酸水平上高度同源,仅吐根堡奶山羊的编码区有2个碱基与其他5种山羊不同,导致编码的两个氨基酸残基也发生了改变。原位杂交结果显示,KAP6-1.2 mRNA在10月份皮肤、胚胎125d皮肤、胚胎115d皮肤初级和次级毛囊的皮质层均有强烈的表达。结论:KAP6-1.2的核苷酸序列和氨基酸序列在不同地区、不同品种山羊中高度保守,在胚胎和成年山羊皮肤的初级和次级毛囊的皮质层均有强烈的表达。Objective： To study the differences of KAP6 - 1.2 for different kinds of goats, and to detect the expression of KAP6 - 1.2 mRNA on the adult and fetal skins of arbas cashmere goat. Methods： This study according to the conservative sequences of announced arbas cashmere goat gene designed a pair of primers of gKAP6 - 1.2,then amplified KAP6- 1.2 cDNA of other kinds of goats, the products were cloned into PMD- 19T vector and sequenced. Made KAP6- 1.2 cRNA probe, In situ hybridization with the adult and fetal skins of arbas cashmere goat. Results： Sequence analysis revealed that nucleofides and encode amino acid sequence were fairly high degree of conservation. Only Toggenburg Goat have two nucleotides and amino acids different from others. In situ hybridization with the KAP6 - 1.2 cRNA probe on skin revealed a strong positive signal in both primary and secondary hair follicles. Conclusion：The nucleofides and encode amino acid sequence of KAP6 - 1.2 were fairly high degree of conservation for different kinds of goats . KAP6 - 1.2 mRNA expressed in cortex of both primary and secondary hair follicles.

关 键 词：绒山羊 角蛋白关联蛋白 高甘氨酸/酪氨酸 原位杂交 

分 类 号：Q785[生物学—分子生物学]