摘要
以3种不同盐度处理的大叶藻为材料,采用RNA转录本5’末端转换机制(SMART)构建了大叶藻叶片全长cDNA文库,原始文库滴度为8.675×106pfu/mL,重组率为97.19%,插入片段平均长度大于800bp,扩增后的文库滴度达到1.03×109pfu/mL。将部分原始λ噬菌体cDNA文库转化成质粒cDNA文库,PCR检测结果显示文库插入片段集中在750～2000bp之间。随机选取20个单克隆进行测序,其中13条序列包含完整的开放阅读框。结果表明所构建文库容量大,全长比例高,为深入开展大叶藻功能基因研究奠定了基础。
The full-length cDNA library of the Z.marina,which were treated with three different salinities,was constructed by adopting the switching mechanism at 5＇end of the RNA transcript（SMART）.The titers of the primary cDNA library and the amplified library were 8.675？？06pfu/mL and 1.03？？09 pfu/mL,respectively.The percentage of recombinant clones was 97.19% and the average length of inserted cDNA fragments was above 800bp.The primary？？hage cDNA library was partially transformed into plasmid cDNA library.Fifty clones were randomly selected for screening the size of the inserted cDNA fragments through PCR and the results showed that the size was around 750？？2000bp.Twenty randomly selected cDNA clones were sequenced and thirteen sequences included completed ORF.The results showed that the library had a large number of clones and a high ratio of full-length cDNA,which will lay a foundation for further functional genomic research in Z.marina.
出处
《中国海洋大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第11期85-89,共5页
Periodical of Ocean University of China
基金
高等学校博士点专项科研基金新教师课题(20070423027)
山东省自然科学基金项目(Q2007E02)
海洋公益性行业科研专项项目(200805075)资助
关键词
大叶藻
全长CDNA文库
耐盐基因
Zostera marina full-length cDNA library salt resistant gene
