摘要
为了获得国家二级保护植物品种菰的功能基因信息,并克隆功能基因片段,研究以SMART(switching mechanism at 5 end of RNA transcript)方式合成菰全长cDNA,结合DSN(duplex-specific nuclease)均一化技术,构建菰叶片组织的均一化全长cDNA文库。该cDNA文库库容量大,达到了3.6×10~6 PFU/m L,插入片段平均长度为1 000 bp,重组率为97.9%。均一化全长cDNA文库能有效富集低丰度表达基因,降低冗余率,适用于目的基因的筛选,以及后续的基因、蛋白互作分析。
In order to obtain functional genes,a narmalized stems cDNA library was constructed from precious plant Zizania latifolia(Griseb.)Stapf.SMART(switching mechanism at5end of RNA transcript)cDNA synthesis combined with DSN(duplex-specific,nuclease)normalization was applied to construct the normalized full-length cDNA library of Z.latifolia.The titer of cDNA library was about3.6×106PFU/mL and the average insertion size was about1.0kb with high recom bination rate(97.9%).Homogenization full-lengthcDNA library enrichment of low abundance is suitable for the purpose of gene selection and subsequent analysis of the gene and proteininteractions,for expressing genes effectively and reducing the redundancy rate.
作者
臧剑
颜育民
张志刚
邹世湘
彭琼
ZANG Jian1;YAN Yu-min;ZHANG Zhi-gang;ZOU Shi-xiang;PENG Qiong(Guangdong Gudao Agricultural Limited Company, Guangzhou 511400, PRC;Hunan Hybrid Rice Research Center,Changsha 410125, PRC;Hunan Wangxianglong Agricultural Limited Company, Changsha 410125, PRC;Hunan Agricultural Biotechnology Research Center, Changsha 410125, PRC)
出处
《湖南农业科学》
2017年第4期11-13,18,共4页
Hunan Agricultural Sciences
基金
湖南农业科技创新资金(2016QN08)
关键词
菰
均一化
CDNA
文库
功能基因
Zizania latifolia
normalization
cDNA library
functional gene
作者简介
臧剑（1972-），男，湖南沅江市人，工程师，主要从事水稻育种研究。;通讯作者
