上西早生柿ETR5基因片段的克隆与植物表达载体的构建  

Cloning of ETR5 from Uenishiwase Persimmon Fruits and Its Plant Expression Vector Construction

作　　者：陈佳[1] 马俊莲[1] 张子德[1] 唐霞[1] 

机构地区：[1]河北农业大学食品科技学院,河北保定071001

出　　处：《华北农学报》2007年第4期25-28,共4页Acta Agriculturae Boreali-Sinica

基　　金：河北省自然科学基金项目(C2006000465)

摘　　要：从上西早生(Uenishiwase)柿叶片中,提取了基因组DNA。以基因组DNA为模板,经PCR扩增,克隆到一个549 bp的ETR5的DNA片段。序列分析表明,上西早生柿基因与洋梨ETR5的核苷酸同源性为79.0%,氨基酸同源性为81.0%;并且含有其他植物ETR5的保守氨基酸区和不变氨基酸残基,表明克隆到的序列是上西早生柿ETR5的基因片段。设计了2对带限制性内切酶位点的特异性引物,以测序质粒为模板,PCR扩增到2个ETR5-Uenishiwase片段。2个片段经单酶切消化后连接成反向互补的大片断。连接片断经双酶切消化后,连接到植物表达载体pBI221上,构建成Uenishiwase-ETR5基因的植物表达载体。One fragment of ETR..5 was cloned from Uenishiwase persimmon by PCR. Sequence analysis indicated that nucleotide identities of Uenishiwase with Pyrus communis was 79%, and amino acid identities were 81%. Uenishiwase has the conserved amino acid regions and invariant amino acid residues of ETR..5 existing in other plants, the cloned production was Uenishiwase persimmon ETR..5 gene sequence confirmly. Two pairs of primers containing restriction enzyme site were designed and were used to amplify sequenced plasmid. Two PCR products were digested by the corresponding restricted enzymes respectively, then connect them to be a long segment that is reverse complemented. Finally inserted it into the expression vector pBI221 after digesting.

关 键 词：柿 上西早生 乙烯受体 ETR5 植物表达载体 

分 类 号：S665.203[农业科学—果树学]