关键词: 辽宁绒山羊; 毛角蛋白Ⅱ型中间丝基因; 毛囊特异性表达载体; 转基因体细胞

Abstract: The objective of this study was to construct a follicle-specific vector expression of keratin type Ⅱ intermediate filament 9(KIFⅡ-9),then transfer it into Liaoning Cashmere goat fetal fibroblasts cells to obtain a transgenic cell by selection,which could be used for nuclear transfer. KIFⅡ-9 cDNA was amplified by RT-PCR from Xinji Fine Wool sheep. Then pcDNA3.1-KK,a follicle-specific expression vector of KIFⅡ-9,was constructed by connecting with KAP6-1 gene promoter sequence amplified by PCR from Xinji Fine Wool sheep and,KIFⅡ-9 cDNA. The Liaoning Cashmere goat fetal fibroblast cells were transfected with the expression vector by lipofectamine,and the transgenic cell clones were obtained after selecting by G418. These results have paved the way to obtain the new transgenic cashmere goat strain by nuclear transfer in the future.

Key words: Liaoning Cashmere goat; keratin type Ⅱ intermediate filament gene; follicle-specific expression vector; transgenic somatic cell 