AUTHOR=Han Xiaohong , Huang Chenyan , Qi Huan , Zhu Yukun , Hu Xinran , Wen Yingxin , Long Yirong , Xu Lei , Zhang Feiping 

TITLE=The construction and evaluation of secretory expression engineering bacteria for the trans-Cry3Aa-T-HasA fusion protein against the Monochamus alternatus vector

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2024.1362961

DOI=10.3389/fcimb.2024.1362961

ISSN=2235-2988

ABSTRACT=<p>Pine wood nematode disease is currently the most deadly forest disease in China, and the <italic>Monochamus alternatus</italic> is its primary vector. Controlling the <italic>M. alternatus</italic> is crucial for managing pine wood nematode disease. This study, based on the selected HasA (pGHKW4) secretory expression vector, used electroporation to combine the genetically modified high-toxicity toxin Cry3Aa-T with the entomopathogenic bacterium <italic>Yersinia entomophaga</italic> isolated from the gut of the <italic>M. alternatus</italic>. The SDS-PAGE and Western blotting techniques were employed to confirm the toxin protein’s secretion capability. The engineered bacteria’s genetic stability and effectiveness in controlling <italic>M. alternatus</italic> were assessed for their insecticidal activity. The results of the SDS-PAGE and Western blotting analyses indicate that the HasA system effectively expresses toxin protein secretion, demonstrates certain genetic stability, and exhibits high insecticidal activity against <italic>M. alternatus.</italic> This study constructed a highly toxic entomopathogenic engineered bacterial strain against <italic>M. alternatus</italic> larvae, which holds significant implications for controlling <italic>M. alternatus</italic>, laying the foundation for subsequent research and application of this strain.</p>