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 - 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 activates expression of other vir genes --> the products 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 genome 2.VirG gene from host genome gets integrated into the bacterial genome 3.T-DNA from host genome gets integrated into the bacterial genome 4.VirG from bacteria gets integrated into the host genome

Identify the correct sequence of steps in T-DNA integration 1.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 excise 3.Plant wound  phenolics  sense by VirG  signal passed to VirA  T-DNA excise 4.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-Plasmid 2.A simple plasmid DNA with expression promoter can be used 3.A plasmid with T-DNA in needed for host genome integration 4.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 selection 2.Three antibiotic resistance genes: one for agro, one for E. coli, and another for plant 3.One antibiotic resistance gene: one for agro selection 4.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 removal 2.Plant cells resulting from enzymatic breakdown of protoplasm 3.Plant cells resulting from enzymatic breakdown of cell wall 4.Plant cells resulting from the enlarged vacuole

Identify correct statement: during Plasmid DNA isolation, EDTA is added In the P1 buffer 1.To inactivate RNAse 2.To inactivate DNAse 3.To inactivate proteases 4.To inactivate cellulases

During Plasmid DNA isolation, the separation of bacterial chromosomal And plasmid DNA occurs as 1.Chromosomal DNA stays in the supernatant, the plasmid DNA precipitates 2.Both DNA stay in the supernatant 3.Chromosomal DNA precipitates while the plasmid DNA stays in the supernatant 4.Both DNA precipitates with different affinities

Identify the correct statement: During PCR of NUD7 gene, the forward and reverse primers represent 1.The start of and the end of the gene sequence respectively 2.The end of and the start of the gene sequences respectively 3.The start of and the End of the plasmid DNA respectively 4.The middle section of and the start of the NUD7 gene sequence respectively