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NEETZoologyBiotechnology - Principles and Processes

A biotechnologist wants to use Agrobacterium tumefaciens to deliver a desirable gene into a dicot plant. To ensure that the plant does not develop crown gall disease after the gene transfer, which specific modification must be made to the wild-type Ti plasmid?

Options

  1. AThe disease-causing genes must be removed to disarm the plasmid while keeping the transfer mechanism intact
  2. BThe entire T-DNA region must be deleted to prevent the integration of any bacterial DNA into the plant genome
  3. CThe origin of replication (ori) must be excised to stop the plasmid from multiplying inside the plant cells
  4. DAntibiotic resistance genes must be added to actively suppress the tumor-inducing property of the bacteria

Correct answer

A. The disease-causing genes must be removed to disarm the plasmid while keeping the transfer mechanism intact

Step-by-step solution

The correct modification is to disarm the Ti plasmid by removing its disease-causing (tumor-inducing) genes. In nature, the T-DNA of the Ti plasmid carries genes that transform normal plant cells into tumor cells. To use it as a safe cloning vector, these pathogenic genes are removed while the sequences required for DNA transfer are kept intact. The gene of interest is then inserted into this disarmed T-DNA. Deleting the entire T-DNA would prevent any gene transfer. Excising the origin of replication would stop the

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