Green fluorescent protein as a marker for expression of a second gene in transgenic plants

被引:91
作者
Harper, BK [1 ]
Mabon, SA [1 ]
Leffel, SM [1 ]
Halfhill, MD [1 ]
Richards, HA [1 ]
Moyer, KA [1 ]
Stewart, CN [1 ]
机构
[1] Univ N Carolina, Dept Biol, Greensboro, NC 27402 USA
基金
美国国家科学基金会;
关键词
green fluorescent protein; transgene presence/expression; transgenic plants; Bacillus thuringiensis cry1Ac;
D O I
10.1038/15114
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:1125 / 1129
页数:5
相关论文
共 26 条
[1]  
Allen GC, 1996, PLANT CELL, V8, P899, DOI 10.1105/tpc.8.5.899
[2]   HIGH-EFFICIENCY TRANSFORMATION OF CULTURED TOBACCO CELLS [J].
AN, GH .
PLANT PHYSIOLOGY, 1985, 79 (02) :568-570
[3]   Gene flow from transgenic crops [J].
Chevre, AM ;
Eber, F ;
Baranger, A ;
Renard, M .
NATURE, 1997, 389 (6654) :924-924
[4]   Engineered GFP as a vital reporter in plants [J].
Chiu, WL ;
Niwa, Y ;
Zeng, W ;
Hirano, T ;
Kobayashi, H ;
Sheen, J .
CURRENT BIOLOGY, 1996, 6 (03) :325-330
[5]   Green-fluorescent protein fusions for efficient characterization of nuclear targeting [J].
Grebenok, RJ ;
Pierson, E ;
Lambert, GM ;
Gong, FC ;
Afonso, CL ;
HaldemanCahill, R ;
Carrington, JC ;
Galbraith, DW .
PLANT JOURNAL, 1997, 11 (03) :573-586
[6]   Codon usage limitation in the expression of HIV-1 envelope glycoprotein [J].
Haas, J ;
Park, EC ;
Seed, B .
CURRENT BIOLOGY, 1996, 6 (03) :315-324
[7]   Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly [J].
Haseloff, J ;
Siemering, KR ;
Prasher, DC ;
Hodge, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (06) :2122-2127
[8]  
HELM R, 1996, CURR BIOL, V8, P178
[9]   A SIMPLE AND GENERAL-METHOD FOR TRANSFERRING GENES INTO PLANTS [J].
HORSCH, RB ;
FRY, JE ;
HOFFMANN, NL ;
EICHHOLTZ, D ;
ROGERS, SG ;
FRALEY, RT .
SCIENCE, 1985, 227 (4691) :1229-1231
[10]   SPONTANEOUS HYBRIDIZATION BETWEEN OILSEED RAPE (BRASSICA-NAPUS) AND WEEDY BRASSICA-CAMPESTRIS (BRASSICACEAE) - A RISE OF GROWING GENETICALLY-MODIFIED OILSEED RAPE [J].
JORGENSEN, RB ;
ANDERSEN, B .
AMERICAN JOURNAL OF BOTANY, 1994, 81 (12) :1620-1626