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苦荞BAC文库的构建及BAC末端序列分析
  • ISSN号:1001-4454
  • 期刊名称:中药材
  • 时间:0
  • 页码:-
  • 分类:R282.71[医药卫生—中药学;医药卫生—中医学]
  • 作者机构:[1]贵州师范大学荞麦产业技术研究中心,贵州贵阳550001, [2]中国中医科学院中药研究所,北京100700
  • 相关基金:国家自然科学基金(31471562,31171609); 国家现代农业产业技术体系专项资金(CARS-08-A4); 贵州省高层次人才培养计划(黔科合人才【2015】4020号); 贵州省荞麦工程技术研究中心(黔科合农G字【2015】4003号)
  • 相关项目:荞麦属植物种子蛋白亚基基因序列及其在染色体上的分布规律研究
中文摘要:

目的:为了深入挖掘苦荞的基因组信息,筛选与苦荞品质相关的功能基因。方法:以苦荞品种"晋荞麦2号"为材料,参考Peterson等方法通过提取高分子量核DNA,经酶切、连接和转化构建苦荞基因组BAC文库;随机挑选48个BAC克隆进行序列末端测序。结果:该文库包括30 000个克隆,插入片段平均大小约为123 kb,空载率小于1%,覆盖苦荞基因组6.9倍。共得到89条BAC末端序列,通过序列比对分析发现,7条(8%)有比对结果。其中,有末端序列与rpoTm2、Trrap、MDR1等已知基因相似,这些基因与DNA锚定,物质跨膜运输和磷酸转移酶活性等功能有关。分析还发现末端序列40G19-F可能是苦荞基因组的重复序列。结论:苦荞BAC文库构建为今后苦荞基因的克隆及全基因组测序奠定了基础。

英文摘要:

Objective: To further study Fagopyrum tataricum genome and to screen the functional genes. Methods: The variety JINQIAO No. 2( Fagopyrum tataricum) native to Shanxi,was used for BAC library construction. The high molecular weight DNA( HMWDNA) was isolated from Fagopyrum tataricum leaves used the methods of Peterson. The HMW-DNA was cut by Hind Ⅲ and ligated to BAC vectors; the ligations were transformed into Escherichia coli DH10 B. Then 48 BAC clones were randomly selected and sequenced BAC end sequences. Results: The library consisted of 30 000 clones with an average insert size of about 123 kb and the empty clone ratio was less than 1%. The library represented an equivalent about 6. 9 fold size of Fagopyrum tataricum genome. Which obtained 89 BAC end sequences,among which 7 sequences( 8%) had alignment results when comparing with NCBI Gen Bank database. The blast hits contained some genes with known function,such as rpo Tm2,Trrap and MDR1 genes,etc. Those genes were associated with DNA binding,transmembrane transport and phosphotransferase activity function. And it was also found that the sequence of 40G19-F was probably repetitive sequences in Fagopyrum tataricum genome. Conclusion: The BAC library will be helpful for the gene cloning and whole genome sequencing of Fagopyrum tataricum.

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期刊信息
  • 《中药材》
  • 中国科技核心期刊
  • 主管单位:国家食品药品监督管理局
  • 主办单位:国家食品药品监督管理局中药材信息中心站
  • 主编:元四辉
  • 地址:广州市中山二路24号中粤大厦10楼
  • 邮编:510080
  • 邮箱:
  • 电话:020-81888465 81889570 87665465
  • 国际标准刊号:ISSN:1001-4454
  • 国内统一刊号:ISSN:44-1286/R
  • 邮发代号:
  • 获奖情况:
  • 全国医药信息成果二等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰文摘与引文数据库,美国生物医学检索系统,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:54900