论文部分内容阅读
以‘戴多星’黄瓜为试材,应用同源克隆和RACE技术从茎尖中得到赤霉素合成的关键酶——贝壳杉烯氧化酶(KO)基因cDNA全长序列,命名为CKO,GenBank登录号为JN792591,其长度为1 895 bp,开放阅读框(ORF)编码519个氨基酸,相对分子量为59.211 kD,等电点(PI)8.45。氨基酸同源性分析发现,CKO与苦瓜的KO同源性最高,达84%;序列结构分析表明,CKO属于细胞色素超家族P450系,具有细胞色素P450的血红素结构域FXXGXRXCXG和跨膜结构域。实时荧光定量分析表明,CKO在遮荫的徒长苗中表达量最高,是正常苗的5.18倍,在遮阴条件下喷施多效唑可降低期表达。
The full-length cDNA of kaolinite oxidase (KO), a key enzyme involved in gibberellin synthesis, was obtained from stem tips by using homologous cloning and RACE technology. The GenBank accession number is JN792591, which is 1 895 bp in length. The open reading frame (ORF) encodes 519 amino acids with a relative molecular mass of 59.211 kD and an isoelectric point (PI) of 8.45. Amino acid homology analysis showed that the KO homology between CKO and bitter gourd was the highest (84%). Sequence analysis showed that CKO belonged to the cytochrome superfamily P450, the heme domain FXXGXRXCXG with cytochrome P450 and the transmembrane domain . The results of real-time fluorescence quantitative analysis showed that the expression of CKO was the highest in shady legumes, which was 5.18 times higher than that of normal seedlings. Under the condition of shading, paclobutrazol could reduce the expression of CKO.