Genetic Transformation of Plants: Methods and Approaches to Develop Transgenic plants

-A source of foreign DNA containing the desired gene

-A vector that carries the gene

-Means to introduce the vector into host plant

Steps in producing Transgenic

- Agrobacterium mediated transformation- Electroporation- Biolistic methods for direct entry- Protoplast fusion

Methods to create Transgenics

Agrobacterium mediated Transgenic

ANIMATIONS REMOVED

Plant wound --> phenolics --> sense by VirA--> signal passed to VirG --> VirG activatesexpression of other vir genes --> theproducts of vir genes help to excise T-DNA from the Ti-plamsid

Steps in T-DNA integration

Identify the right statement: during Agrobacterium mediated transformation

1. T-DNA from bacterial genome gets integrated into the host genome2. VirG gene from host genome gets integrated into the bacterial genome3. T-DNA from host genome gets integrated into the bacterial genome4. VirG from bacteria gets integrated into the host genome

Identify the correct sequence of steps in T-DNA integration1. Plant wound phenolics sense by VirA signal passed to VirG T-DNA excise 2. Phenolics plant wound sense by VirA signal passed to VirG T-DNA excise3. Plant wound phenolics sense by VirG signal passed to VirA T-DNA excise4. Plant wound Signal passed to VirG phenolics sense by VirA T-DNA excise

Identify the correct statement: For gene gun method of transformation

1. you need bacteria with Ti-Plasmid2. A simple plasmid DNA with expression promoter can be used3. A plasmid with T-DNA in needed for host genome integration4. No –TDNA is required

Identify the correct statement: The agrobacterium based plasmid should Maintain

1. Two antibiotic resistance genes: one for agro selection and for plant selection2. Three antibiotic resistance genes: one for agro, one for E. coli, and another for plant3. One antibiotic resistance gene: one for agro selection4. Two antibiotic resistance gene: one for E. coli selection and another for plant selection

Identify the correct statement: Protoplast refers to

1. Plant cells resulting from plasma membrane removal2. Plant cells resulting from enzymatic breakdown of protoplasm3. Plant cells resulting from enzymatic breakdown of cell wall4. Plant cells resulting from the enlarged vacuole

Identify correct statement: during Plasmid DNA isolation, EDTA is addedIn the P1 buffer

1. To inactivate RNAse2. To inactivate DNAse3. To inactivate proteases4. To inactivate cellulases

During Plasmid DNA isolation, the separation of bacterial chromosomalAnd plasmid DNA occurs as

1. Chromosomal DNA stays in the supernatant, the plasmid DNA precipitates2. Both DNA stay in the supernatant3. Chromosomal DNA precipitates while the plasmid DNA stays in the supernatant4. Both DNA precipitates with different affinities

Identify the correct statement: During PCR of NUD7 gene, the forward and reverse primers represent1. The start of and the end of the gene sequence respectively2. The end of and the start of the gene sequences respectively3. The start of and the End of the plasmid DNA respectively4. The middle section of and the start of the NUD7 gene sequence respectively