Using “Reverse Genetics” to Learn the Biological Function of a Gene

 

Leicht, B. J. & C. Cheng

Tested Studies in Laboratory Teaching, 2010, Volume 31

Abstract

Reverse genetics begins with the DNA sequence of a gene and works toward ascertaining the mutant phenotype. Because its genome is fully sequenced, the plant Arabidopsis thaliana is amenable to reverse genetics. A large collection of T-DNA insertion lines has been generated and sites of insertion within the Arabidopsis genome have been mapped. This workshop describes use of websites to identify T-DNA insertion mutations in a gene of interest and to design PCR primers that identify plants that are homozygous or heterozygous for the insertion. The exercise reinforces concepts in gene segregation through the genotyping of individual plants from insertion lines.

Keywords:  Arabidopsis, reverse genetics, T-DNA

University of Delaware (2009)