Translator Disclaimer
1 July 2000 Tagging Bloodmeals with Phagemids Allows Feeding of Multiple-Sample Arrays to Single Cages of Mosquitoes (Diptera: Culicidae) and the Recovery of Single Recombinant Antibody Fragment Genes from Individual Insects
Author Affiliations +
Abstract

A recombinant single-chain variable-region human antibody fragment (scFv) was expressed inEscherichia coli,extracted in hypertonic sucrose, mixed directly with blood and fed toAnopheles gambiaeGiles mosquitoes. WhenE. colicontaining the phagemids that encode these scFv were included in bloodmeals, phagemids could be recovered from the mosquito midgut for up to 3 d after feeding. Furthermore, large arrays of such gene-tagged scFv-containing bloodmeals could be fed to cages of mosquitoes using microtiter plates. Arrays of phagemids with and without an antibody insert were fed to single cages of mosquitoes to test whether individual mosquitoes fed from single wells of such arrays. Phagemids were recovered from 95% of blood-fed females and >80% of these phagemids were monoclonal. Therefore, it is possible to feed multiple sample arrays of recombinant proteins to single cages of mosquitoes and to recover the genetic material that encodes for only one of the array elements from individual mosquitoes. This demonstration indicates that multiple-sample feeding and recovery strategies are feasible and may represent a viable strategy for future rapid screening of biologically active genes, gene products or microorganisms in live arthropods.

Gerry F. Killeen, Brian D. Foy, Mohammed Shahabuddin, Wendy Roake, Andrew Williams, Tristan J. Vaughan, and John C. Beier "Tagging Bloodmeals with Phagemids Allows Feeding of Multiple-Sample Arrays to Single Cages of Mosquitoes (Diptera: Culicidae) and the Recovery of Single Recombinant Antibody Fragment Genes from Individual Insects," Journal of Medical Entomology 37(4), 528-533, (1 July 2000). https://doi.org/10.1603/0022-2585-37.4.528
Received: 22 September 1999; Accepted: 1 February 2000; Published: 1 July 2000
JOURNAL ARTICLE
6 PAGES

This article is only available to subscribers.
It is not available for individual sale.
+ SAVE TO MY LIBRARY

SHARE
ARTICLE IMPACT
RIGHTS & PERMISSIONS
Get copyright permission
Back to Top