Green fluorescent protein as a marker for expression of a second gene in transgenic plants.
The use of transgenic crops has generated concerns about transgene movement to unintended hosts and the associated ecological consequences. Moreover, the in-field monitoring of transgene expression is of practical concern (e.g., the underexpression of an herbicide tolerance gene in crop plants that are due to be sprayed with herbicide). A solution to these potential problems is to monitor the presence and expression of an agronomically important gene by linking it to a marker gene, such as GFP. Here we show that GFP fluorescence can indicate expression of the Bacillus thuringiensus cry1Ac gene when co-introduced into tobacco and oilseed rape, as demonstrated by insect bioassays and western blot analysis. Furthermore we conducted two seasons of field experiments to characterize the performance of three different GFP genes in transgenic tobacco. The best gene tested was mGFP5er, a mutagenized GFP gene that is targeted to the endoplasmic reticulum. We also demonstrated that host plants synthesizing GFP in the field suffered no fitness costs.
Harper
B K
BK
Department of Biology, University of North Carolina, Greensboro, NC 27402-6174, USA.
Mabon
S A
SA
Leffel
S M
SM
Halfhill
M D
MD
Richards
H A
HA
Moyer
K A
KA
Stewart
C N
CN
D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001427:Bacterial Toxins; D001937:Brassica / Q000235:genetics / Q000378:metabolism*; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006460:Hemolysin Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D010947:Plants, Toxic; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D008164:Luminescent Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D049452:Green Fluorescent Proteins