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

Abstract

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

CLC Number:

Q813

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.

References

1 张文. 内蒙古绒山羊毛被生长规律的研究. 呼和浩特: 内蒙古农业大学,2011.
2 张燕军. 内蒙古绒山羊胎儿期皮肤毛囊形态发生和发育规律及部分Hox基因家族成员研究. 呼和浩特: 内蒙古农业大学,2007.
3 高爱琴. 成纤维生长因子5(FGF5)基因cDNA克隆及在不同品种绵羊和山羊中的多态性研究. 呼和浩特: 内蒙古农业大学,2005.
4 鲍珣,张拓,张涌. 人端粒酶逆转录酶(hTERT)基因对山羊胎儿成纤维细胞的影响 [J].中国畜牧兽医, 2009, 36(8): 42~46.
5 Basilico L, Abbondi M, Fumagalli A. Influence of pertussis toxin on thermic responses to morphine and neurotensin in rats[J].Eur J Pharmacol,1992, 222(2~3):241~245.
6 Drogemuller C, Rufenacht S, Wichert B, et al. Mutations within the FGF5 gene are associated with hair length in cats[J].Anim Genet,2007,38(3):218~221.
7 Eswarakumar V P, Lax I, Schlessinger J. Cellular signaling by fibroblast growth factor receptors[J].Cytokine Growth Factor Rev,2005,16:139~149.
8 Grose R, Dickson C. Fibroblast growth factor signaling in tunorigenesis[J].Cytokine Growth Factor Rev,2005,16:179~186.
9 Hanada K, Perry-Lalley D M, Ohnmacht G A, et al. Identification of fibroblast growth factor-5 as an over expressed antigen in multiple human adenocarcinomas[J].Cancer Res,2001,61:5511~5516.
10 Herbert J M, Rosenquist T, Martin G R. FGF5 as a regulator of the hair growth cycle: Evidence from targeted and spontaneous mutations[J].Cell,1994,78:1017~1025.
11 Housley D J, Venta P J. The Long and the short of it: Evidence that FGF5 is a major deter minant of canine 'hair’-itability[J]. Anim Cenet, 2006,37(4):309~315.
12 Kehler J S, David V A, Schaffer A A, et al. Four independent mutation in the feline fibroblast growth factor 5 gene determine the long-haired phenotype in domestic cats[J].J Hered,2007,98(6):555~566.
13 Li C X, Jiang M S, Chen S Y, et al. Correlation analysis between single nucleotide polymorphism of FGF5 gene and wool yield in rabbits[J].Yi Chuan,2008,30(7):893~899.
14 Mansour S L, Thomas K R, Capecchi M R. Disruption of the proto-oncogene int-2 in mouse embryo-drived stem cells: A general strategy for targeting mutations to non-selectable genes[J].Nature,1988,336(6197):348~352.
15 Rossant J, Ciruna B, Partanen J. FGF signaling in mouse gastrulation and anteroposterior patterning[J].Cold Spring Harb Symp Quant Biol,1997,62:127~133.
16 Zhan X, Bates B, Hu X G, et al. The human FGF-5 oncogene encodes a novel protein related to fibroblast growth factors[J].Mol Cell Biol,1988,8:3487~3495.

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

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