[目的]克隆浙贝母环阿屯醇合成酶(Cycloartenol synthase,CAS)5`末端cDNA序列。[方法]利用简并引物及RT-PCR方法获得CAS的核心基因,应用cDNA末端快速扩增(Rapid-amplification of cDNA ends,RACE)技术快速克隆5`末端片段,采用NCBI网站上...[目的]克隆浙贝母环阿屯醇合成酶(Cycloartenol synthase,CAS)5`末端cDNA序列。[方法]利用简并引物及RT-PCR方法获得CAS的核心基因,应用cDNA末端快速扩增(Rapid-amplification of cDNA ends,RACE)技术快速克隆5`末端片段,采用NCBI网站上的Protein Blast软件进行氨基酸同源性比对。[结果]从浙贝母叶片中成功地克隆出一条长691bp的CAS基因5`末端cDNA片段,含起始密码子及163bp的5`延伸区。对5`末端的150个氨基酸同源性分析表明,该片段与C.asiatica,G.glabra,A.thaliana,W.somnifera的CAS同源性分别达72%,66%,71%,71%。[结论]上述结果表明所克隆的片段为浙贝母CAS的5`末端序列,为下一步CAS全长基因的克隆及CAS在浙贝母次生代谢中的功能研究打下基础。展开更多
The 5’ end cDNA fragment(ER1’) of early responsive gene(ER1) of rice(Oryza sativa L. subsp. japonica No.4) induced by blast fungus M. grisea has been isolated and sequenced (606 bp) by 5’RACE technique(Fig. 2). DNA...The 5’ end cDNA fragment(ER1’) of early responsive gene(ER1) of rice(Oryza sativa L. subsp. japonica No.4) induced by blast fungus M. grisea has been isolated and sequenced (606 bp) by 5’RACE technique(Fig. 2). DNA sequence analysis showed that ER1’ (606 bp) can encode 134 amino acids and there are 309 bp nucleotides located at the 3’ end of the untranslated region of mRNA of ER1 gene(Fig. 3). The 89 bp overlapping fragment was found between the 5’ end of ER1 cDNA fragment and the 3’ end of ER1’(Fig. 3). The partial identities of amino acid sequence deduced from ER1’ with an unknown protein(gene accession number AC002409) of Arabidopsis thaliana was 82%(Fig. 4).展开更多
文摘The 5’ end cDNA fragment(ER1’) of early responsive gene(ER1) of rice(Oryza sativa L. subsp. japonica No.4) induced by blast fungus M. grisea has been isolated and sequenced (606 bp) by 5’RACE technique(Fig. 2). DNA sequence analysis showed that ER1’ (606 bp) can encode 134 amino acids and there are 309 bp nucleotides located at the 3’ end of the untranslated region of mRNA of ER1 gene(Fig. 3). The 89 bp overlapping fragment was found between the 5’ end of ER1 cDNA fragment and the 3’ end of ER1’(Fig. 3). The partial identities of amino acid sequence deduced from ER1’ with an unknown protein(gene accession number AC002409) of Arabidopsis thaliana was 82%(Fig. 4).
