复叶槭FtsZ基因cDNA片段的克隆及RNA干扰载体的构建

Cloning of FtsZ gene cDNA fragment of Acer negundo L.and constructon of RNAi vectors

利用简并引物RT-PCR,首次从复叶槭中克隆到569 bp的FtsZ基因cDNA片段,其与GenBank已经登录的FtsZ基因氨基酸序列的同源性为96.90%,并具有FtsZ蛋白的核心功能序列GGGTGSG。从上述序列中选取300 bp的序列,分别克隆其正、反义基因片段,同时,克隆GUS基因的1个内含子,作为间隔区基因。将正义、间隔区和反义3个基因片段串联,插入到植物表达载体p3300-35ST中,构建复叶槭RNA干扰载体p35ST-AnFtsZi。为接下来遗传转化、培育复叶槭彩叶新品种奠定基础。

By degenerate primers negundo L. for the first time. It is GenBank, and it contains FtsZ gene selected from the above sequence and the same time, an intron was cloned RT-PCR, a 569 bp fragment of FtsZ gene cDNA was acquired from Acer 96.90% homological with FtsZ gene amino acid sequence registered in family＇s function center sequence GGGTGSG. A 300 bp sequence was cloned to get its sense fragment and anti-sense fragment, respectively. At from GUS gene as the spacer fragment. Then linked the 3 fragments of sense, anti-sense and spacer, inserted to plant expression vector p3300-35ST, constructed a RNA interference plant expression vector p35ST-AnFtsZi. The experiment makes preparation for the genetic transforming and Acer negundo L. ornamental breeding.