摘要
为构建绒山羊FGF5基因敲除载体及探明FGF5基因更多的序列信息,通过PCR方法分别扩增获得FGF5基因上游启动子至第一外显子1.6 kb片段、第一内含子至部分第二外显子8.0 kb片段及1.8 kb、2.4 kb和4.5 kb三段缺口片段。分别对以上PCR扩增片段进行测序,通过序列比对分析拼接获得了全长为9483 bp FGF5基因的部分序列,其结构包括编码区上游1 255 bp、第一外显子364 bp,第一内含子7 809 bp、部分第二外显子55 bp。并对绒山羊、绵羊、牛的FGF5基因启动子序列进行了分析,结果显示绒山羊和绵羊只有5个核苷酸的差异,而与牛的序列比较发现差异较大,牛的启动子中缺失了一个24 bp的DNA序列。本研究获得了绒山羊FGF5基因启动子至第二外显子部分序列的共9 483 bp的序列,为构建绒山羊FGF5基因敲除载体及探究FGF5基因的功能奠定了基础。
This experiment was conducted to obtain the FGF5 gene fragment with range from promoter to the partial fragment of the second exon. Arbas Cashmere goat as experiment material,several DNA fragments from FGF5 gene were obtained by PCR amplification as follows： the 1. 6 kb fragment including promoter and the first exon,the 8. 0 kb fragment including partial first exon,first intron and partial second exon and three gap DNA fragments： 1. 8 kb,2. 4 kb and 4. 5 kb. All PCR products above were sequenced and all sequences were assembled into a complete sequence of 9483 bp,and exon 1 is 364 bp,intron 1 is 7809 bp,partial the exon 2 is55 bp. Moreover,the promoters of FGF5 gene in Cashmere goat,sheep,bovine were analyzed. Results showed that five differences in promoter region between Cashmere goat and sheep,and a DNA fragment with 24 bp was missing in promoter region in bovine comparing with Cashmere goat and sheep. This goat FGF5 sequence with 9483 bp was obtained successfully,which will provide the help for constructing the target vector and functional research of FGF5 gene.
出处
《内蒙古农业大学学报（自然科学版）》
CAS
北大核心
2014年第6期60-67,共8页
Journal of Inner Mongolia Agricultural University(Natural Science Edition)
基金
国家自然科学基金地区重点项目(30960244)
转基因生物新品种培育重大专项(2008ZX08008-002)
内蒙古自然科学基金重大项目(2013ZD06)
