摘要： 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(Short hairpin RNA)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。
关键词:
草莓,
FaEtr2基因,
RNA干扰,
遗传转化
Abstract: In order to delay the ripening and softening process of strawberry,RNA interference technology was used to inhibit strawberry ethylene receptor gene expression. Two pairs of primers containing restriction enzyme sites were designed and used to amplify sequenced plasmid. PCR products and the plasmid pBI121 were digested by the corresponding restricted enzymes respectively,and linked directionally. Then the shRNA expression vector can be designed. After transcription,the hairpin mRNA obtained due to the inverted-repeat DNA fragment. The constructed expression vector was transformed into Agrobacterium LBA4404 for the following-up genetic transformation research. Four strains of transgenic plants were detected by PCR and GUS histochemical staining.
Key words:
Strawberry,
FaEtr2 gene,
RNAi,
Genetic transformation
中图分类号:
宋春丽, 马俊莲, 唐霞, 张子德, 赵丛枝, 李静. RNA干扰技术抑制草莓FaEtr2基因表达的研究[J]. 华北农学报, 2014, 29(1): 28-30. doi: 10.7668/hbnxb.2014.01.006.
SONG Chun-li, MA Jun-lian, TANG Xia, ZHANG Zi-de, ZHAO Cong-zhi, LI Jing. Inhibition of Strawberry FaEtr2 Gene Expression by RNA Interfering[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(1): 28-30. doi: 10.7668/hbnxb.2014.01.006.