Functional Genomics: Using Reverse Genetics to Test Bioinformatics Predictions

In this project, students (i) amplify by PCR and clone an internal portion of a selected gene from Agrobacterium tumefaciens, (ii) use restriction enzymes and gel electrophoresis to verify that they have cloned the correct DNA fragment, (iii) transform the cloned DNA fragment into wildtype A. tumefaciens and select for crossover recombinants in which the gene of interest is disrupted, and (iv) use the mutant A. tumefaciens strain to test the bioinformatics-predicted function of the gene of interest. The focus is on student involvement in original research.