转抗菌肽基因鱼腥草的筛选及稳定性分析

Screening of Houttuynia cordata Thunb. Transformed with Foreign Antimicrobial Peptide and Analysis of Its Stability

【目的】筛选转基因鱼腥草,并初步分析外源基因在鱼腥草植株中的稳定性。【方法】以选择培养基中生长的转基因鱼腥草为材料,通过卡那霉素(Kan)叶片涂抹法进一步筛选抗性植株,并进行Kan抗性基因nptⅡ和目的基因CN(cecropin B/NP-1)的PCR分析。【结果】Kan叶片涂抹法筛选的最佳浓度为5 000 mg/L,观察时间以4 d为宜;经PCR检测验证,Kan抗性植株的nptⅡ基因阳性率达到96%,其中目的基因CN阳性率为37%。【结论】建立了适用于转基因鱼腥草的快速筛选方法,转基因鱼腥草中目的基因CN的稳定性低于抗性基因。

Objective To screen transgenic Houttuynia cordata Thunb.and to analyze the stability of exogenous genes initially.Methods Kanamycin was smeared to the surface of the leaves to screen the transgenic plants containing the npt Ⅱ gene.And then PCR detection of the npt Ⅱ gene and target gene CN（cecropin B/NP-1）was carried out in kanamycin-resistance plant.Results The results of kanamycin-smearing screening method showed that the optimal kanamycin concentration was 5000 mg/L and the optimal observation time was day 1 to day 4.The result of PCR analysis showed that the expression ratio of resistant gene npt Ⅱ was 96% in the resistant plants,and the expression ratio of target gene CN was 37%.Conclusion A method for quickly screening transgenic Houttuynia cordata Thunb.has been established.The stability of target gene in the transgenic plant is inferior to the selecting maker gene