学术
极大节旋藻（Arthrospiramaxima）高分子量基因组文库构建及其应用
茅云翔[1] 凌娜[1] 王广策[2] 隋正红[1] 张学成[1]
[1]中国海洋大学海洋生命学院,青岛266003 [2]中国科学院海洋研究所,青岛266071
摘 要:
构建了极大节旋藻fosmid文库。该文库含有4300个克隆，重组率是100％，插入片段集中在30～36kh之间，平均为33kb，覆盖率为节旋藻基因组的25．8倍。随机挑选100个克隆进行双末端测序，得到110个序列，经生物信息学分析筛查到67条功能基因序列。选取了14对特异的末端序列作为序列标签位点并设计引物，采用STS-PCR反应池法通过多轮筛选，得到127个阳性克隆，按照相互位置关系，绘制了4个连续的克隆重叠群。该研究为深入了解极大节旋藻分子遗传特性和绘制物理图谱奠定了基础。[著者文摘]
文章出处:
《高技术通讯》-2007年17卷2期 -191-196页
栏目信息:
分 类 号:
Construction of high molecular weight genome library of Arthrospira maxima and its application
Mao Yunxiang, Ling Na, Wang Guangce , Sui Zhenghong, Zhang Xuecheng （College of Marine Life Sciences, Ocean University of China, Qingdao 266003; Institute of Oceanology, Chinese Academy of Scienses, Qingdao 266071 ）
Abstract:
The fosmid library of Arthrospira maxima was constructed. It contains 4300 clones, and the recombination frequency is 100%. The most insert fragments are between 30 kb and 36 kb and the average insertion length is 33 kb. As a result,the library is 25.8 genome-equivalent coverage. One hundred clones were selected randomly to dual-ends sequence and 110 sequences were obtained. 67 functional genes were screened based on biological infonnatics analysis. Fourteen spe-cial end sequences were chosen as the sequence site-tagged sites and designed primers. 127 positive clones were obtained through several screenings by STS-PCR pooling protocol. According to their position relationships, four continuous contigs were protracted. This study will lay the foundations for further understanding of molecular genetics features and drawing the physical map of Arthrospira.[著者文摘]
Key words:
Arthrospira maxima, fosmid library, sequence-tagged site, functional genes, contig, physical map
基金资助:
国家自然科学基金（30200208,30571418）与中心科学院海洋研究所实验海洋生物学重点实验室资助
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