【摘要】 缺刻缘绿藻(Myrmecia incise)富含花生四烯酸(arachidonic acid,AA),是迄今为止所知道的花生四烯酸含量最高的藻类。采用Trizol法提取总RNA,然后按照Clontech公司的SMART cDNA文库构建的方法,构建了缺刻缘绿藻的cDNA文库,测得的原始文库滴度为6×106pfu/mL,随机挑选文库的阳性单克隆进行PCR鉴定,扩增出的片段主要集中在0.5~1.5kb,平均插入片段长度约为1kb,文库的重组率达95.83%,说明本实验所构建的cDNA文库质量合格。更多还原

【Abstract】 The freshwater green microalga Myrmecia incisa was isolated from Mt.Tateyama,Japan.It is well-known by far that this microalga is the richest source of one polyunsaturated fatty acid(PUFA),arachidonic acid(20:4ω6,AA).Total RNA was isolated from this alga by Trizol method,and the cDNA library of M.incisa was constructed using SMART cDNA Library Construction Kit,Clontech,Inc.The titer of unampified library reached 6×106 pfu/mL,and the recombination rate was about 95.83%.PCR amplification of randomly selected positive clones illustrated the inserted cDNA fragments ranged from 0.5 kb to 1.5 kb,and the average length was about 1 kb.All these results suggested that the cDNA library was successfully constructed.更多还原