1 / 15

T-DNA Mutagenesis

T-DNA Mutagenesis . T ransfer-DNA Mutagenesis: a chemical or physical treatment that creates changes in DNA sequence which can lead to mutation strains that are passed on to the next generation. -PURPOSE:

saniya
Download Presentation

T-DNA Mutagenesis

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. T-DNA Mutagenesis

  2. Transfer-DNA Mutagenesis: a chemical or physical treatment that creates changes in DNA sequence which can lead to mutation strains that are passed on to the next generation.

  3. -PURPOSE: To create loss of function mutations in order to determine the function of a gene.

  4. T-DNA Flanking Region RB T-DNA LB KAN Pdi2 -HOW DOES THIS WORK: Vector transmission by way of Agro plant is randomly inserted into the nuclei chromosomal sites.

  5. -Agro? Agrobacteria tumefaciens is a bacteria found on certain plants that were found to cause tumors on wounded plant areas. Found to contain Ti (Tumor inducing) plasmid that creates a mutation in the plants genomic sequence. The Ti plasmid’s ability to integrate itself into a DNA sequence was isolated and the tumor inducing quality was taken out.

  6. -LOSS OF FUNCTION MUTATIONS: is when a mutation is created in such a way that death does not occur so as to observe the effects on the plant by the loss of a certain gene. In other words, a gene is knocked out and the plant is grown and observed for any differences between the mutant strain and the control strain. Thus facilitating (understanding) the function of that knocked-out gene.

  7. Pdi2 A LB T-DNA Primer F1 Wild Type R1 primer T-DNA in the Arabidopsis Genome T-DNA – from Ti plasmid in the Agrobacteria tumefaciens.. Uses the insertional quality to carry foreign genes into the plant genome.

  8. T-DNA Flanking Region RB T-DNA LB KAN Pdi2 -VALID TEST RESULTS: To achieve valid data from the gel results. Homozygous cells must be used, not Heterozygous. Genotype - Segregation

  9. -HOMOZYGOUS: 2 of the same (genes). On gel, homozygous will only produce one band on either the wild type control side or the T-DNA control side, not both. Needed for accurate results.

  10. -HETEROZYGOUS: different genes. On gel, will produce a band on both the WT control side and the T-DNA control side. Not used for verification of T-DNA.

  11. -GEL ELECTROPHORESIS: 1. Verify if sample is homozygous, 2. Verify that T-DNA knockout is not there, 3. Verify that T-DNA is there and inserted. WT A1 A2 TDNA+ - WT+ A1 A2 WT - WT A1 A2 WT+ - WT A1 A2 TDNA+ -

  12. WT A1 A2 WT+ - WT A1 A2 TDNA+ -

  13. WT A1 A2 TDNA+ - WT A1 A2 WT+ -

  14. Pdi2 A LB T-DNA Primer F1 Wild Type R1 primer -disrupted genes do not make RNA. That function has been knocked out. -F1 and R2 primer will make WT. -F1 and LB primer will make T-DNA knockout.

  15. Future Advances in Biotechnology Mutagenesis Research: BEFORE AFTER

More Related