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RNA干扰技术抑制草莓FaEtr2基因表达的研究

DOI: 10.7668/hbnxb.2014.01.006, PP. 28-30

Keywords: 草莓,FaEtr2基因,RNA干扰,遗传转化

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Abstract:

采用RNA干扰技术抑制草莓乙烯受体基因表达,以延缓草莓成熟软化进程。以测序质粒为模板,PCR扩增草莓FaEtr2基因片段。双酶切正、反义PCR产物及表达载体,将酶切产物定向连接到pBI121载体上,构建成该基因的shRNA(ShorthairpinRNA)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,可应用于RNA干扰的研究。将构建好的载体转化根癌农杆菌LBA4404,用基因工程菌菌液浸染千代田草莓组培苗叶片,用卡那霉素筛选抗性植株。PCR检测和GUS组织化学染色检测到4个株系的转基因植株。

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