Steps of Creating a transgenic plant in lab

Creating a transgenic plant involves several steps in the laboratory using modern biotechnology techniques. Below is an overview of the process to create a transgenic plant:-

1. 1. Identify the Desired Gene: The first step is to identify and isolate the gene with the desired trait that you want to introduce into the target plant. This gene could come from another plant species, animals, bacteria, or any other source.

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3. 2. Prepare the Gene of Interest: Once the desired gene is identified, it needs to be isolated and amplified through techniques like polymerase chain reaction (PCR) to create multiple copies of the gene.

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5. 3. Construct the Plant Expression Vector: The isolated gene is then inserted into a plant expression vector. A plant expression vector is a small circular piece of DNA that acts as a carrier to transfer the gene into the plant cells. The vector contains regulatory sequences that control the expression of the gene, ensuring it functions correctly in the plant.

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7. 4. Introduce the Vector into Plant Cells: The constructed plant expression vector is now introduced into plant cells. There are various methods to achieve this, including:

   a. Agrobacterium-mediated transformation: Using a soil bacterium called Agrobacterium tumefaciens, the vector is transferred into the plant cells naturally infected by the bacterium.

   b. Biolistic (or gene gun) method: Tiny gold or tungsten particles coated with the plant expression vector are shot into plant cells using a gene gun.

   c. Electroporation: Applying a brief electric shock to the plant cells to create temporary pores through which the vector can enter.

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9. 5. Regenerate Transgenic Plants: Once the plant cells have taken up the vector and integrated the foreign gene into their genome, they are grown on a special medium to develop into transgenic plantlets. The plantlets are then transferred to the soil to grow into mature transgenic plants.

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11. 6. Screening and Selection: Not all plant cells will successfully incorporate the foreign gene. Therefore, the transformed plant cells need to be identified and selected from the non-transformed ones. Selective markers (e.g., antibiotic or herbicide resistance genes) are often included in the plant expression vector to aid in this process. Only the transformed cells will survive in the presence of the marker.

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13. 7. Confirmation of Transgene Integration: The presence and stability of the transgene in the regenerated plants need to be confirmed. Techniques like PCR and Southern blotting are used to verify the presence and copy number of the introduced gene.

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15. 8. Testing and Characterization: The transgenic plants with confirmed gene integration are subjected to extensive testing and characterization to assess the expression of the desired trait and to ensure there are no unintended effects.

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17. 9. Field Trials and Regulatory Approval: Before commercial use, transgenic plants usually undergo field trials to assess their performance and potential environmental impact. Regulatory authorities review the safety and efficacy data to grant approval for commercial cultivation.

It is crucial to mention that creating transgenic plants requires specialized laboratory facilities, skilled researchers, and adherence to strict biosafety protocols and regulatory guidelines. Moreover, potential environmental and ethical concerns should be addressed during the development and commercialization of transgenic plants