Transformation of internal extracellular bacteria isolated from Rhagoletis completa cresson gut with enhanced green fluorescent protein.
We discovered Zeocintrade mark is an effective antibiotic against Enterobacter agglomerans and Klebsiella pneumoniae strains isolated from the walnut husk fly (Rhagoletis completa Cresson: Family Tephritidae) and that bleomycin resistance can be used as a selective marker in transforming plasmids. We transformed Ent. agglomerans and K. pneumoniae strains originally isolated from their close association with R. completa gut to produce enhanced green fluorescent protein, a variant of green fluorescent protein in the first demonstration of genetic transformation of internal extracellular bacteria isolated from a tephritid pest. We report methods for plasmid-mediated transformation of these bacteria, the expression of fluorescent marker protein from the transforming plasmids, and the stability of the transforming plasmid in the bacteria. We also discuss applications of this technology in the study of pest biology and control implementation.
Peloquin
J J
JJ
Department of Entomology, University of California, Riverside, CA 92521, USA.
Kuzina
L
L
Lauzon
C R
CR
Miller
T A
TA
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001761:Bleomycin / Q000494:pharmacology; D004175:Diptera / Q000382:microbiology*; D004352:Drug Resistance, Microbial / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D018274:Electroporation; D004755:Enterobacteriaceae / Q000187:drug effects / Q000235:genetics*; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D010957:Plasmids / Q000032:analysis; D013997:Time Factors; D014169:Transformation, Bacterial
D000900:Anti-Bacterial Agents; D005819:Genetic Markers; D008164:Luminescent Proteins; D001761:Bleomycin; D049452:Green Fluorescent Proteins