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Assessment of genetically engineered Trabulsiella odontotermitis as a ‘Trojan Horse’ for paratransgenesis in termites

机译:评估转基因的沙门氏菌牙白质白蚁作为“特洛伊木马”用于白蚁的超转基因

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Background The Formosan subterranean termite, Coptotermes formosanus is an invasive urban pest in the Southeastern USA. Paratransgenesis using a microbe expressed lytic peptide that targets the termite gut protozoa is currently being developed for the control of Formosan subterranean termites. In this study, we evaluated Trabulsiella odontotermitis , a termite-specific bacterium, for its potential to serve as a ‘Trojan Horse’ for expression of gene products in termite colonies. Results We engineered two strains of T. odontotermitis , one transformed with a constitutively expressed GFP plasmid and the other engineered at the chromosome with a Kanamycin resistant gene using a non- disruptive Tn7 transposon. Both strains were fed to termites from three different colonies. Fluorescent microscopy confirmed that T. odontotermitis expressed GFP in the gut and formed a biofilm in the termite hindgut. However, GFP producing bacteria could not be isolated from the termite gut after 2?weeks. The feeding experiment with the chromosomally engineered strain demonstrated that T. odontotermitis was maintained in the termite gut for at least 21?days, irrespective of the termite colony. The bacteria persisted in two termite colonies for at least 36?days post feeding. The experiment also confirmed the horizontal transfer of T. odontotermitis amongst nest mates. Conclusion Overall, we conclude that T. odontotermitis can serve as a ‘Trojan Horse’ for spreading gene products in termite colonies. This study provided proof of concept and laid the foundation for the future development of genetically engineered termite gut bacteria for paratransgenesis based termite control.
机译:背景技术台湾地下白蚁,Coptotermes formosanus是美国东南部的一种入侵性城市害虫。目前正在开发使用针对白蚁肠道原生动物的微生物表达的裂解肽进行副转基因,以控制台湾地下白蚁。在这项研究中,我们评估了白蚁特异性细菌Trabulsiella odontotermitis作为白蚁菌落中表达基因产物的“特洛伊木马”的潜力。结果我们设计了两种菌株的牙周炎性牙菌,一种是用组成型表达的GFP质粒转化的,另一种是使用卡那霉素抗性基因在染色体上使用了非破坏性的Tn7转座子进行了工程改造的。两种菌株都被喂食了来自三个不同菌落的白蚁。荧光显微镜证实,牙周炎舌头炎在肠中表达GFP,并在白蚁后肠中形成生物膜。但是,在2周后不能从白蚁肠中分离出产生GFP的细菌。用染色体工程菌株进行的饲养实验表明,无论白蚁菌落如何,在白蚁肠中都可以保持牙本质T.牙龈炎至少21天。喂食后,细菌在两个白蚁菌落中持续存在至少36天。该实验还证实了在伴侣之间T. Odontotermitis的水平转移。结论总的来说,我们得出结论,弓形体牙周炎可以作为“特洛伊木马”在白蚁菌落中传播基因产物。这项研究提供了概念验证,并为基于遗传工程的白蚁肠道细菌的未来发展奠定了基础。

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