论文部分内容阅读
目的:将抗菌肽cecropin B和兔NP-1(CN)融合基因转入鱼腥草,培育鱼腥草基因工程新品种。方法:设计并合成抗菌肽融合基因CN,构建重组质粒pBI121-CN,并转入农杆菌LBA4404,以农杆菌介导法转化鱼腥草外植体,将卡那霉素筛选获得的再生抗性植株以快速筛选PCR法进一步筛选阳性植株,再提取基因组DNA进行PCR-Southern鉴定,RT-PCR分析目的基因在鱼腥草中的表达,以大肠杆菌K12抑菌圈实验和立枯丝核菌感染实验分析转基因鱼腥草抗菌能力。结果:抗菌肽基因已整合到转基因鱼腥草基因组中并表达,表现出抗菌能力增强的性状。结论:以抗菌肽融合基因CN转化鱼腥草,初步获得抗菌活性提高的转基因植株。
OBJECTIVE: To transfer the antibacterial peptide cecropin B and rabbit NP-1 (CN) fusion gene into Houttuynia cordata and cultivate a new gene engineering variety of Houttuynia cordata. METHODS: The antibacterial peptide fusion gene CN was designed and synthesized. The recombinant plasmid pBI121-CN was constructed and transferred into Agrobacterium LBA4404. The Houttuynia cordata explants were transformed by Agrobacterium tumefaciens and regenerated resistance was obtained by screening with kanamycin. Plants were further screened for positive plants by rapid screening PCR, followed by extraction of genomic DNA for PCR-Southern analysis, RT-PCR analysis of the expression of the target gene in Houttuynia cordata, E. coli K12 inhibition zone and Rhizoctonia solani infection. The experiment analyzed the antibacterial ability of the transgenic Houttuynia cordata. Results: The antibacterial peptide gene has been integrated into the genome of the transgenic Houttuynia cordata and expressed, showing an enhanced antibacterial activity. Conclusion: Transgenic H. pylori was transformed with antibacterial peptide fusion gene CN, and transgenic plants with improved anti-bacterial activity were obtained.