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蜘蛛基因组DNA Cosmid文库构建和拖丝蛋白基因的克隆

李敏;DavidL.Kaplan

  The genomic DNA Cosmid library was constructed fromNephila clavipesspider muscle using SuperCos 1 Cosmid as a vector.The title of library was >5×104 cfu/μg ligated DNA.On the basis of published sequence from a partial cDNA sequence of the 3′end of the dragline silk gene, we designed and synthesized 3 oligonucleotides.Oligonucleotides were labeled with nonradioactive digoxigenindUTP and detected with chemilluminescent substrate.56 positive recombinants were screen from the Cosmid library using DIGOligo 2 as a probe.DNA dot hybridization using DIGOligo 1 and DIGOligo 3 as the probes, respectively, 3 positive signals were identified from 56 colonies.They were appeared the same pattern when DNA from the colones digested by restriction enzymes.The spider dragline silk gene was confirmed again by Southern blot hybridization.……   
[关键词]:蜘蛛;基因组文库;拖丝;丝蛋白基因
[文献类型]:期刊
[文献出处]: 《动物学报2001年06期
[格式]:PDF原版; EPUB自适应版(需下载客户端)