应用重组PCR技术构建大豆脂肪氧化酶基因RNA干扰表达载体

Use Recombiant PCR to Construct RNAi Expression Vector of Soyabean Lipoxygenase

RNA干扰是近年来生物技术领域新兴的研究热点之一。在作物的研究中,RNA干扰被认为在作物品质改良中具有重要的应用价值,并已在水稻、小麦、玉米等作物中得到成功应用,但在大豆的研究中尚鲜有报道。在研究植物RNA干扰的实验中,构建相应的RNA干扰表达载体是常用手段,但传统构建方法较费时费力。重组PCR是一种利用重叠延伸的方法进行高效和快速的体外基因片段拼接的PCR技术。研究将重组PCR技术应用于构建大豆脂肪氧化酶基因ihpRNA干扰表达载体,一方面探索一条更简便、快捷、适应范围更广泛的大豆基因RNA干扰表达载体构建方法,另一方面所构建的载体为进一步研究RNA干扰在大豆脂肪氧化酶改良中的应用奠定了基础。结果显示应用重组PCR技术构建大豆脂肪氧化酶基因RNA干扰表达载体是可行的,相比与常规方法此方法更简便、快捷、效率更高。

For the past few years RNAi was a newly rising territory in biotechnology. RNAi was believed to have huge potential in quality improvement of crops, and it had been successfully applied in quality improvement of rice, wheat and maize, while its application in soybean quality improvement was less documented. During the research of RNAi in plant, RNAi expression vector is always be used, and the commonly used method is time and labor consuming. Recombinant PCR is a high performance method of gene splicing by overlapping. In this research ,we use recombinant PCR to construct RNAi expression vector of soybean lipoxygenase ,in order to find a easier, faster and generally way to construct RNAi expression vector for soybean. Results showed that constructing RNAi expression vector of the lipoxygenase gene by recombinant PCR technology is feasible, and the method is more rapid, simple and convenient than common methods.