摘要： 分析克隆的柿果ACC合成酶基因的序列，根据其与表达载体pSMAK311上的酶切位点设计2对带酶切位点的特异性引物，以测序质粒为模板，PCR扩增柿果ACC合成酶基因片段。双酶切PCR回收产物以及表达载体，将酶切产物定向连接构建成反义表达载体；在此基础上构建该酶基因的shRNA表达载体，由此转录的mRNA因两端序列反向互补而成发夹式RNA，因此，可应用于RNA干扰研究。将所构建好的载体转化农杆菌EHA101用于后续的遗传转化研究。

关键词: 柿果, ACC合成酶, RNA干扰, 植物表达载体

Abstract: According to the restriction enzyme site of the expression vector and cloning sequence, two pairs of primers containing restriction enzyme site were desigined and used to amplify sequenced plasmid. PCR products and the plasmid pSMAK311 were digested by the corresponding restricted enzymes respectively, and linked directionally. Then the shRNA expression vector can be designed. After sequencing transcription, the hairpin mRNA obtained due to the inverted-repeat DNA fragment. The constructed expression vector was transformed into Agrobacterium EHA101 for the following-up research.

Key words: Persimmon, ACC synthase, RNA interference, Plant expression vector

中图分类号: 

• S665.2