Studies On Arbas Cashmere Goat FGF5Gene Knockout

Mutations of FGF5are able to lead to extension of hair follicle;growth phase and significant increase.of hair length in kinds of mammals, promoting that the cashmere yield of Arbas cashmere goat can be raised by inhibiting the expression FGF5.In this study, transgenic cell line was established by gene knockout, and used as nuclear donor for somatic cell nuclear transfer aimed to product gene knockout cloning cashmere goats.Firstly, the obtained Arbas cashmere goat fetal skin fibroblast cells by primary culture of organization adherence method grow in good condition, and can be used for gene targeting and somatic cell nuclear transfer.Secondly, through positive and negative selection51cell clones were got after transfection, and finally cryopreserved12clones via following propagagation. On DNA level,5positive clones were acquired after PCR amplification, and on RNA level,4positive were indentified ultimately after RT-PCR. Compared with normal fibroblast, tremendous changes have taken place in the FGF5gene-knockouted positive cell clones in cell morphology, traits and growth characteristics:bigger celll volume; fuzzy boundaries, reduced stereoscopic impression; growth rate dramatic decline, significant increase of trypsin digestion time.Used transgenic positive cells as nuclear donor and matured goat oocytes in vitro as nuclear receptor for somatic cell nuclear transfer, the formation reconstructed embryos developed to the blastocyt stage. The cleavage reconstructed embryos transplanted into39receptor goats, which were failed to produce any transgenic somatic cell cloned goat.It suggests that the variations in cell morphology and growth characteristics might be affect by transfection, drug selection, long-term culture in vitro, or relating to FGF5gene knockout; the failure to produce a transgenic somatic cell cloned individual may be associated with the donor cells, or FGF5gene knockout had an influence on development of reconstructed embryos. This study laid a foundation in developing high cashmere yield Arbas cashmere goats by molecular breeding methods.