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用单个特异引物和通用引物ｏｌｉｇｏ（ｄＴ）１５从桃受伤叶片ｃＤＮＡ中扩增出１．３ｋｂ左右大小的片段．将该扩增产物克隆并对重组克隆作酶切分析发现至少可分为３类．用桃ＡＣＣ氧化酶基因组ＤＮＡ为探针进行点杂交表明，４，７，９，１０，１１，１２重组质粒能与之杂交，其中７，９，１０，１１，１２号重组质粒酶切图谱与已报导的桃果实成熟相关的ＡＣＣ氧化酶ｃＤＮＡ基因基本一致，而４号克隆则不同．ＤＮＡ序列分析和ＰＣＲ鉴定表明，插入片段均由５′端特异引物单个引物扩增出来，对７号重组克隆插入片段ＤＮＡ全序列分析表明，７号重组克隆插入片段全长１１４６ｂｐ，包含了除启始密码子外的全部编码区和１９２ｂｐ的３′端非编码区．通过补上起始密码子ＡＴＧ构建重组表达载体，在大肠杆菌中表达出３５×１０３的非融合蛋白
A single specific primer and a universal primer oligo (dT) 15 were used to amplify a fragment of about 1.3 kb from the peach injured leaf cDNA. Cloning of the amplified product and digestion analysis of the recombinant clones revealed that it could be divided into at least three categories. Dot hybridization using peach ACC oxidase genomic DNA as a probe showed that 4, 7, 9, 10, 11, 12 recombinant plasmids could hybridize with them, and the restriction maps of 7, 9, 10, 11, Reported peach fruit ripe ACC oxidase cDNA genes are basically the same, while the No. 4 clone is different. DNA sequence analysis and PCR identification showed that the inserted fragments were all amplified by a single primer specific to 5 ’end. The complete DNA sequence analysis of recombinant clone 7 showed that the full length of the recombinant clone 7 was 1146bp, The entire coding region outside the start codon and the 3 ’non-coding region of 192 bp. The recombinant expression vector was constructed by adding the start codon ATG, and 35 × 103 non-fusion protein was expressed in E. coli