Abstract
For the screening of transfer DNA (T-DNA) integration in transgenic plant material, we developed a method based on specific amplification of genomic plant DNA flanking T-DNA borders. This approach is possible because the length of the region flanking T-DNA extremity on a restriction fragment is specific to the integration locus. We have modified an adaptor ligation PCR technique developed for amplification of unknown DNA flanking known sequence. The PCR patterns obtained were specific and reproducible for different plants from a given transgenic line. Furthermore, the number of PCR products obtained could be considered a good estimation of the T-DNA copy number. When compared to Southern blot analysis, the PCR results give valuable complementary information about the complexity of the T-DNA integration pattern and also about the integrity of the T-DNA borders. We describe the applications of this approach to populations of transgenic Arabidopsis thaliana plants.
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Claims 3
Claim · model-written summary
PCR patterns were specific and reproducible for plants from a given transgenic line.
- Taxon
- Arabidopsis thaliana
Arabidopsis thalianae1 - Outcome
- PCR patternse0
- Value
- specific and reproduciblee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The PCR patterns obtained were specific and reproducible for different plants from a given transgenic line.”
e1“transgenic Arabidopsis thaliana plants”
Claim · model-written summary
PCR product number provided a good estimate of T-DNA copy number.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Outcome
- estimation of the T-DNA copy numbere0
- Value
- goode0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the number of PCR products obtained could be considered a good estimation of the T-DNA copy number”
e1“transgenic Arabidopsis thaliana plants”
Claim · model-written summary
Compared with Southern blot analysis, PCR supplied complementary information on T-DNA integration complexity and border integrity.
- Taxon
- Arabidopsis thalianaArabidopsis thalianae1
- Outcome
- complexity of the T-DNA integration pattern and also about the integrity of the T-DNA borderse0
- Comparator
- Southern blot analysise0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“When compared to Southern blot analysis, the PCR results give valuable complementary information about the complexity of the T-DNA integration pattern and also about the integrity of the T-DNA borders.”
e1“transgenic Arabidopsis thaliana plants”