香石竹ACC氧化酶基因的克隆及其正义反义表达载体的构建

ACC Oxidase Gene Clonging in Carnation and the Construction of Its' Plant Expression Vectors and Antisense Expression Vectors

利用在Genebank上已报道的香石竹ACC氧化酶(ACO)基因的cDNA序列设计特异引物,以香石竹品种‘MASTER’的cDNA为模板进行PCR扩增,克隆香石竹ACC氧化酶(ACO)基因。测序结果显示,克隆基因的序列与报道序列完全一致。将克隆的ACC氧化酶(ACO)基因分别连接到植物表达载体PBI121启动子CaMV 35S的上游及下游,构建香石竹ACC氧化酶(ACO)基因的正义表达载体PBI121-ACO及反义表达载体PBI121-anti ACO。经PCR鉴定,基因已成功构建到表达载体上。为香石竹抗衰老基因工程育种奠定了基础。

Basing on the eDNA sequence of ACC Oxidase Gene from carnation in Genebank, we designed a pair of special primers , according to PCR, amplified ACC Oxidase Gene of carnation using the cDNA of MASTER as template. The sequence analysis showed that the cloned sequence was accord with the reported totally, PCR product was cloned into the upstream and downstream of CaMV 35S promoter in plant expression vector-PBI121 plasmid separately, established expression vectors（PBI121-ACO） and antisense expression vectors（PBI121-antiACO）. PCR identification showed gene had been cloned into expression vector successfully which can be used for old resistant breeding of gene engineering.