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从油桃‘中油四号’中克隆了3个CYP707A家族成员(PpCYP707A1、PpCYP707A2和PpCYP707A3)的全长,测序发现其CDS序列与桃基因组公布的序列一致。同源比对发现CYP707A家族在不同物种间具有高度的序列保守性,并且都含有CYP707A家族基因比较保守的血红素铁配位体半胱氨酸残基;系统进化树分析表明,PpCYP707A1与马铃薯CYP707A1、花生CYP707A1、菜豆CYP707A1、菜豆CYP707A2的亲缘关系较近,桃CYP707A2与拟南芥CYP707A2的亲缘关系较近,桃CYP707A3与拟南芥CYP707A4、花生CYP707A2的亲缘关系较近。荧光定量PCR分析表明,PpCYP707A1在茎中的表达量较高,PpCYP707A2在叶中的表达量较高,PpCYP707A3在花中的表达量较高,并且桃CYP707A家族一般在成熟组织中的表达水平要高于幼嫩组织。推测桃CYP707A家族成员交叉调控花芽休眠进程,并且发现花芽中的PpCYP707A2可以被ABA和GA诱导。原核诱导并纯化PpCYP707A1和PpCYP707A2蛋白可以为后续蛋白的功能鉴定奠定基础。
Three CYP707A family members (PpCYP707A1, PpCYP707A2 and PpCYP707A3) were cloned from nectarine ’Zhongyou 4’ and sequenced. The sequence of CDS was consistent with the sequence published in peach genome. Homology comparison revealed that the CYP707A family is highly conserved among different species and contains a conserved heme iron cysteine residue with a CYP707A family of genes. Phylogenetic tree analysis showed that PpCYP707A1 and CYP707A1 , Peanut CYP707A1, kidney bean CYP707A1 and kidney bean CYP707A2. The genetic relationship between CYP707A2 and CYP707A2 in Arabidopsis thaliana was close, and the genetic relationship between CYP707A3 and CYP707A2 in Arabidopsis was closer. Fluorescence quantitative PCR analysis showed that the expression level of PpCYP707A1 in the stem was higher, the expression level of PpCYP707A2 in leaves was higher, the expression level of PpCYP707A3 in flowers was higher, and the expression level of CYP707A family was generally higher in mature tissues In young organizations. It is speculated that peach CYP707A family members cross-regulate flower bud dormancy, and found that PpCYP707A2 in flower buds can be induced by ABA and GA. Prokaryotic induction and purification of PpCYP707A1 and PpCYP707A2 proteins could lay the foundation for the subsequent functional identification of proteins.