敲除成纤维细胞生长因子5基因对阿尔巴斯绒山羊胎儿皮肤成纤维细胞系的影响

›› 2012, Vol. 39 ›› Issue (10): 27-32.

敲除成纤维细胞生长因子5基因对阿尔巴斯绒山羊胎儿皮肤成纤维细胞系的影响

高蓓¹, 马玉珍^2,3, 尹俊^4,5, 周欢敏¹

1. 内蒙古农业大学,动物生物技术重点实验室,内蒙古呼和浩特 010018;
2. 内蒙古大学,哺乳动物繁殖生物学与生物技术重点实验室,内蒙古呼和浩特 010021;
3. 内蒙古自治区医院,内蒙古呼和浩特 010017;
4. 内蒙古农业大学生命科学学院,内蒙古呼和浩特 010018;
5. 内蒙古大学生命科学学院,内蒙古呼和浩特 010021

收稿日期:2012-03-23 出版日期:2012-10-20 发布日期:2012-10-19
通讯作者: 周欢敏(1959-),男,教授,博士生导师,研究方向:动物遗传育种与胚胎工程。E-mail:huanminzhou@263.net E-mail:huanminzhou@263.net
作者简介:高蓓(1986-),女,内蒙古人,硕士生,研究方向:动物遗传与发育。
基金资助:
高产绒量转基因绒山羊新品种培育(2008ZX08008-002)。

Influences of FGF5 Gene Knockout to Arbas Cashmere Goat Fetal Skin Fibroblast Cell Line

GAO Pei¹, MA Yu-zhen^2,3, YIN Jun^4,5, ZHOU Huan-min¹

1. Key Laboratory of Animal Biotechnology, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Key Laboratory of Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010021, China;
3. Inner Mongolia Hospital, Hohhot 010017, China;
4. College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China;
5. College of Life Sciences, Inner Mongolia University, Hohhot 010021, China

Received:2012-03-23 Online:2012-10-20 Published:2012-10-19

摘要/Abstract

摘要： 本研究为体细胞核移植生产转基因动物提供核供体,以阳离子脂质体介导转染同源重组成纤维细胞生长因子5(fibroblast growth factor 5,FGF5)基因打靶载体至体细胞中,经正负双筛选获得了51个细胞克隆,扩繁后剩余12个克隆,最终经DNA及RNA水平的分子生物学鉴定得到4个阳性细胞克隆,即建立了敲除FGF5基因的内蒙古阿尔巴斯绒山羊胎儿皮肤成纤维细胞系。与普通细胞相比,所得的阳性细胞克隆在细胞形态、性状及生长特性等方面均发生了极其显著的变化:细胞体积增大;边界模糊,立体感降低;生长速率明显下降;胰蛋白酶消化所需时间明显增加。研究结果表明上述变化可能是受到了敲除FGF5基因、细胞转染和药物筛选作用等的影响所致。

关键词: 基因敲除; FGF5基因; 药物筛选; 细胞形态; 细胞生长特性

Abstract: In this study, in order to provide a nuclear for somatic cell nuclear transfer and production of transgenic animals, cationic liposome-mediated transfection of homologous recombination FGF5 gene knockout vector to somatic cell. After positive and negative selection obtained 51 cell clones, which remaining 12 clones after propagation, and ultimately get 4 positive cell clones by the molecular indentification of DNA and RNA levels, namely established the FGF5-knockouted Inner Mongolia Arbas cashmere goat fetal skin fibroblast cell line. Compared with normal somatic cell, the four FGF5 -knockouted positive cell clones had occurred very significant changes in cell morphology, traits and growth characteristics: bigger cell volume;fuzzy boundaries, reduced stereoscopic;growth rate dramatic decline;trypsin digestion time required was significantly increased. This study showed that these changes might be due to FGF5 gene knockout, cell transfection, drug selection and other roles.

Key words: geneknock; FGF5 gene; drug selection; cell morphology; cell growth characteristics

中图分类号:

Q813

高蓓, 马玉珍, 尹俊, 周欢敏. 敲除成纤维细胞生长因子5基因对阿尔巴斯绒山羊胎儿皮肤成纤维细胞系的影响[J]. , 2012, 39(10): 27-32.

GAO Pei, MA Yu-zhen, YIN Jun, ZHOU Huan-min. Influences of FGF5 Gene Knockout to Arbas Cashmere Goat Fetal Skin Fibroblast Cell Line[J]. , 2012, 39(10): 27-32.

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