Making a Cry1Ab expressing plant: Recombinant DNA technology

I. Engineering plants to express cry proteins

A. The steps

II. Getting the cry1Ab gene from Bt

A. Where is the cry1Ab gene?

B. Genomes

C. Genbank

D. PCR

III. Introducing the Cry1Ab gene into a plant

A. Using Agrobacterium tumefaciens to introduce Cry1Ab into a plant chromosome

B. Generation of a GM plant

Study Questions:

1. How is a bacterial chromosome different from one of your chromosomes? How is it similar?

2. Why is it essential to know the Cry1Ab DNA sequence in order to PCR amplify its DNA sequence?

3. What is PCR? Describe and draw out what happens during the three steps (separation of strands, binding of primers, replication of DNA) that occur every cycle.

4. Would you expect to find the Kan^R DNA sequence in a genetically engineered plant generated using our scheme?

5. How does the Kan^R selectable marker in the T-DNA+Cry1Ab plasmid aid in generation of a plant containing Cry1Ab?

6. At which step in the process of creating the Cry1Ab containing plant does the genetic modification of the plant occur (allowing you to call it a GMO)?

Powerpoint below