桃果实中ACC合酶基因克隆及基因沉默载体构建

摘 要：植物中乙烯是一种具有促进果实成熟和衰老的内源激素。ACC合酶是植物乙烯生物合成途径中一个重要的限速酶，沉默ACC合酶基因的表达能减少植物性内源性乙烯的产生。本研究以中华寿桃为研究材料，采用RT-PCR 技术，克隆获得ACC合酶基因。将该基因酶切回收后连接到pTRV-RNA2载体上，转化DH5α，筛选阳性克隆，进行酶切鉴定。测序后与已知序列进行同源性比较，其同源性达到99.6%，表明将ACC合酶基因成功连接到pTRV-RNA2基因沉默载体上。

Cloning of ACC Synthase Gene and Vector Construction of Gene silencing from peach

Abstract: Ethylene, an internal source hormone, affects fruit ripening and senescence in plants. ACC Synthase is one of the rate-limiting enzymes of ethylene biosynthesis in plant. The ACC Synthase gene silencing can down-regulate the product of endogeny ethylene in plant. In this study, ACC Synthase gene was cloned with RT-PCR from peach, and then was digested and inserted into the pTRV-RNA2 vector, and transformed into the E. coli component DH5α. The recombination plasmid was selected and identified by restriction enzyme analysis. Sequence analysis showed that nucleotide of the ACC Synthase gene was 99% identical to the published sequence. The results showed that ACC Synthase gene was rightly cloned into the gene silencing vector pTRV-RNA2.