Engineering blood meal-activated systemic immunity in the yellow fever mosquito, Aedes aegypti Export

@article{citeulike:4648487, abstract = {Progress in molecular genetics makes possible the development of alternative disease control strategies that target the competence of mosquitoes to transmit pathogens. We tested the regulatory region of the () gene of for its ability to express potential antipathogen factors in transgenic mosquitoes. -mediated transformation was used to integrate a 2.1-kb -promoter fragment driving the expression of the () coding region, one of the major insect immune factors. PCR amplification of genomic DNA and Southern blot analyses, carried out through the ninth generation, showed that the transgene insertion was stable. The transgene was strongly activated in the fat body by a blood meal. The mRNA levels reached a maximum at 24-h postblood meal, corresponding to the peak expression time of the endogenous gene. High levels of transgenic defensin were accumulated in the hemolymph of bloodfed female mosquitoes, persisting for 20–22 days after a single blood feeding. Purified transgenic defensin showed antibacterial activity comparable to that of defensin isolated from bacterially challenged control mosquitoes. Thus, we have been able to engineer the genetically stable transgenic mosquito with an element of systemic immunity, which is activated through the blood meal-triggered cascade rather than by infection. This work represents a significant step toward the development of molecular genetic approaches to the control of vector competence in pathogen transmission.}, author = {Kokoza, Vladimir and Ahmed, Abduelaziz and Cho, Wen-Long and Jasinskiene, Nijole and James, Anthony A. and Raikhel, Alexander}, citeulike-article-id = {4648487}, citeulike-linkout-0 = {http://www.pnas.org/content/97/16/9144.full.abstract}, citeulike-linkout-1 = {http://www.pnas.org/content/97/16/9144.full.full.pdf}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/10908672}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=10908672}, day = {1}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {aedes, anti\_malarial\_peptides, antibacterial, blood\_meal, defensin, malaria, methods, mosquito, transgenic, vitellogenin}, month = {August}, number = {16}, pages = {9144--9149}, posted-at = {2009-05-27 21:03:55}, priority = {0}, title = {Engineering blood meal-activated systemic immunity in the yellow fever mosquito, Aedes aegypti}, url = {http://www.pnas.org/content/97/16/9144.full.abstract}, volume = {97}, year = {2000} }

TY - JOUR ID - citeulike:4648487 L3 - citeulike-article-id:4648487 N2 - Progress in molecular genetics makes possible the development of alternative disease control strategies that target the competence of mosquitoes to transmit pathogens. We tested the regulatory region of the () gene of for its ability to express potential antipathogen factors in transgenic mosquitoes. -mediated transformation was used to integrate a 2.1-kb -promoter fragment driving the expression of the () coding region, one of the major insect immune factors. PCR amplification of genomic DNA and Southern blot analyses, carried out through the ninth generation, showed that the transgene insertion was stable. The transgene was strongly activated in the fat body by a blood meal. The mRNA levels reached a maximum at 24-h postblood meal, corresponding to the peak expression time of the endogenous gene. High levels of transgenic defensin were accumulated in the hemolymph of bloodfed female mosquitoes, persisting for 20–22 days after a single blood feeding. Purified transgenic defensin showed antibacterial activity comparable to that of defensin isolated from bacterially challenged control mosquitoes. Thus, we have been able to engineer the genetically stable transgenic mosquito with an element of systemic immunity, which is activated through the blood meal-triggered cascade rather than by infection. This work represents a significant step toward the development of molecular genetic approaches to the control of vector competence in pathogen transmission. IS - 16 JF - Proceedings of the National Academy of Sciences of the United States of America EP - 9149 TI - Engineering blood meal-activated systemic immunity in the yellow fever mosquito, Aedes aegypti VL - 97 SP - 9144 KW - aedes KW - anti_malarial_peptides KW - antibacterial KW - blood_meal KW - defensin KW - malaria KW - methods KW - mosquito KW - transgenic KW - vitellogenin AU - Kokoza, Vladimir AU - Ahmed, Abduelaziz AU - Cho, Wen-Long AU - Jasinskiene, Nijole AU - James, Anthony AU - Raikhel, Alexander PY - 2000/08/01/ UR - http://www.pnas.org/content/97/16/9144.full.abstract ER -