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The Sequecing Analysis of FGF5 Gene from Cashmere Goat and Construction of FGF5 Gene Knock-out Vector

Author: SuShaoFeng
Tutor: YinJun
School: Inner Mongolia Agricultural University
Course: Genetics
Keywords: FGF5 PCR Goat Sheep Polymorphism Knockout vector Knockout
CLC: Q78
Type: Master's thesis
Year: 2011
Downloads: 27
Quote: 0
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Abstract


Fibroblast growth factor 5 (Fibroblast Growth Factor-5 FGF5) expression in the central nervous system and skin, inhibit hair follicles in the skin by the flourishing period to telogen transformation, FGF5 knock inter coat of mice was significantly longer than that of the control. Other animals, such as dogs, cats and rabbits that FGF5 mutation hair length of. Goats FGF5 variability and the hair is the length of the relationship is unclear. For goats FGF5 gene, the following results were obtained: selection of cashmere goats Arbas material by PCR amplification, cloning FGF5 promoter, first exon, first intron, spliced ??into a full-length sequencing 9483bp the sequence, wherein the first exon of 364bp, the first intron is 7809bp. Laid the foundation for the subsequent analysis of polymorphism and knockout vector construction. The the cashmere goats the FGF5 first intron sequences and GenBank personal genome the Blastn found and cashmere goats the FGF5 first intron showed a high degree of synteny homologous sequences, three high homology region, with borne respectively 69%, 70% and 75%, respectively; two low homology region, the first non-homologous regions located between 1545bp-2368bp and 1545bp-2788bp, 6799bp-6860bp second non-homologous regions in between; two parts, compared with the the bovine the FGF5 first intron sequence of the first intron of the gene of the bovine FGF5 than cashmere goat sequence located between 4728bp-4729bp, 274bp and located between 5378bp-5379bp The 1365bp fragment. Direct sequencing of PCR products, combined with the the sheep cDNA sequence has been released Arbas cashmere goats, dairy goats, Mongolia sheep (sheep), fibroblast growth factor 5 (FGF5) 3 gene exon mutation analysis, such as dairy goat and Mongolian sheep heterozygous loci FGF5 gene from the translation initiation codon the sub (ATG) 273 sites in the presence of T and C, cashmere goats for C. The variability is not caused by a change of the the FGF5 amino acid sequence of the goats, cashmere goat and Mongolian sheep four nucleotide mutation. Sequence the FGF5 gene outside of three varieties of exon forecast into the amino acid sequence, sheep and goats from the translation initiation codon the sub (ATG) 161 sites have a different cashmere goats and dairy goats for the A (alanine) codon GCG; Mongolian sheep V (valine) codon GTG. The cashmere goat FGF5 promoter to the first exon of the 1.6kb fragment is inserted into the knockout vector pLox Xba Ⅰ and Cla Ⅰ point the constructed intermediate knockout vector pLox5; then goat the FGF5 first intron 4.5kb fragment is inserted into Not Ⅰ locus pLox5 the screening in line with the direction of insertion of the carrier named pLox5-33, the knockout vector sequencing analysis proved completely in line with the expected design. Laid the foundation obtained for the latter part of the the cashmere goats individuals knockout FGF5 gene.

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