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目的 构建含有微囊藻毒素 (MC)表达相关基因的基因组文库 ,用于分离MC相关基因或鉴别MC表达藻株。方法 使用MC产毒藻株NIES 90 ,获取基因组DNA ,酶切制备片段 ,克隆到质粒 pUC18中 ,转化JM10 9构成基因组文库。 结果 建成容量覆盖蓝绿藻基因组 10倍以上 ,插入片段介于 2 0 0～ 70 0bp的NIES 90基因组文库 ,其中获得至少一段序列具有NIES 90特异性 ,可能为MC相关基因。结论 构建文库成功 ,可用于后续MC特异性基因研究
Objective To construct a genomic library containing genes related to microcystin (MC) expression and to isolate MC-related genes or to identify MC-expressing algal strains. Methods Genomic DNA was obtained from MC toxin producing strain NIES90. The fragment was digested with restriction endonuclease and cloned into plasmid pUC18. The recombinant plasmid was transformed into JM109 to construct a genomic library. Results The NIES 90 genomic library covering more than 10 times the genome of blue-green algae and 200 ~ 70 bp of insertions was constructed. At least one sequence was found to have NIES 90 specificity, which may be MC-related gene. Conclusion The successful construction of the library can be used for subsequent MC-specific gene research