杉木茎段形成层组织cDNA文库的构建

Construction of cDNA Library from Stem Cambium Tissue of Cuninghamia lanceolata(Lam.) Hook

为了分离和克隆与杉木木材形成相关的重要基因,笔者以杉木2～3年生枝条,5～6月份形成层活动期的茎段形成层组织为材料,用表达型噬菌体载体ZAP构建了cDNA文库.文库宿主菌为E. coli XL1-Blue,亚克隆空菌为E. coli XLOLR.试验结果表明:初级文库的库容量达到6.1×105 pfu,扩增文库的滴度达到5.2×109 pfu/mL,重组率达到95%,插入片段大小在0.5～3.0 kb.

In order to isolate target genes for controlling the wood formation and quality components, a cDNA library was constructed from Cuningharnia lanceolata stems cambium tissue with the ZAP express phage vector. The host bacteria is E. coli XL1-Blue, and subclone bacteria is E. coli XLOLR. The compacity of primary library is 6.1 ×10^5 pfu, and the titer of amplified library is 5.2 ×10^9 pfu/mL, in which 95 % of phages were recombinant, the size of fragments inserted rang from 0.5 kb to 3.0 kb.