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פותח על ידי קלירמאש פתרונות בע"מ -
Visual characterization of recombination at FRT-gusA loci in transgenic tobacco mediated by constitutive expression of the native FLP recombinase
Year:
1996
Source of publication :
Theoretical and Applied Genetics
Authors :
גדעוני, דוד
;
.
גלבוע, נחמה
;
.
לשם, ברוך
;
.
Volume :
93
Co-Authors:
Bar, M., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Leshem, B., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Gilboa, N., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Gidoni, D., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Facilitators :
From page:
407
To page:
413
(
Total pages:
7
)
Abstract:
FLP/FRT-mediated site-specific recombination was studied with a recombination-reporter gene system which allows visualization of β- glucuronidase (GUS) expression after site-specific excisional activation of a silent gusA gene. This system was used for characterization of the functional activity of the Saccharomyces cerevisiae native FLP recombinase driven by the cauliflower mosaic virus (CaMV) 35s promoter [linked to the tobacco mosaic virus (TMV) omega translational leader] in mediating site-specific recombination of chromosomal FRT sites in tobacco FLP x FRT-reporter hybrids. Six hybrids were generated from crosses of lines containing either a stably integrated recombination-reporter or a FLP-expression construct. The activated gusA phenotype was specific to hybrid progenies and was not observed in either parental plants or their selfed progenies. Recombination efficiency in whole seedlings was estimated by the percent of radioactivity on a Southern blot which was incorporated into the recombined DNA product. Estimated efficiency mean values for the six crosses ranged from 5.2 to 52.0%. Histochemical analysis in hybrid plants visualized GUS activity with variable chimeric patterns and intensities. Recombination efficiency and GUS expression varied both among and within crosses, while higher recombination efficiency coincided with larger and more intense patterns of GUS activity. These data suggest that recombination is induced randomly during somatic developmental stages and that the pattern and intensity generated in a given plant are affected by factors imposing varibility not only between but also within crosses. Additionally, while recombination in a population of FLP/FRT hybrids may occur in all plants, recombination efficiency may still be low in any given plant. The activity of the native, as compared to a modified, FLP (Kilby et al. 1995) in the activation of transgenic traits in tobacco is discussed.
Note:
Related Files :
Excisional recombination
Native FLP/FRT
Site-specific
tobacco
Transgene activation
β-Glucuronidase
עוד תגיות
תוכן קשור
More details
DOI :
10.1007/s001220050295
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
28757
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:41
Scientific Publication
Visual characterization of recombination at FRT-gusA loci in transgenic tobacco mediated by constitutive expression of the native FLP recombinase
93
Bar, M., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Leshem, B., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Gilboa, N., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Gidoni, D., Department of Plant Genetics, Institute of Field and Garden Crops, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Visual characterization of recombination at FRT-gusA loci in transgenic tobacco mediated by constitutive expression of the native FLP recombinase
FLP/FRT-mediated site-specific recombination was studied with a recombination-reporter gene system which allows visualization of β- glucuronidase (GUS) expression after site-specific excisional activation of a silent gusA gene. This system was used for characterization of the functional activity of the Saccharomyces cerevisiae native FLP recombinase driven by the cauliflower mosaic virus (CaMV) 35s promoter [linked to the tobacco mosaic virus (TMV) omega translational leader] in mediating site-specific recombination of chromosomal FRT sites in tobacco FLP x FRT-reporter hybrids. Six hybrids were generated from crosses of lines containing either a stably integrated recombination-reporter or a FLP-expression construct. The activated gusA phenotype was specific to hybrid progenies and was not observed in either parental plants or their selfed progenies. Recombination efficiency in whole seedlings was estimated by the percent of radioactivity on a Southern blot which was incorporated into the recombined DNA product. Estimated efficiency mean values for the six crosses ranged from 5.2 to 52.0%. Histochemical analysis in hybrid plants visualized GUS activity with variable chimeric patterns and intensities. Recombination efficiency and GUS expression varied both among and within crosses, while higher recombination efficiency coincided with larger and more intense patterns of GUS activity. These data suggest that recombination is induced randomly during somatic developmental stages and that the pattern and intensity generated in a given plant are affected by factors imposing varibility not only between but also within crosses. Additionally, while recombination in a population of FLP/FRT hybrids may occur in all plants, recombination efficiency may still be low in any given plant. The activity of the native, as compared to a modified, FLP (Kilby et al. 1995) in the activation of transgenic traits in tobacco is discussed.
Scientific Publication
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