Verification of the Isolation of the Chalcone Isomerase Gene from Glycine Max

In this paper we report the verified isolation of the gene encoding the flavonoid-biosynthetic enzyme chalcone flavonone isomerase (CHI) from Glycine max as well as the procedures used to generate the results. We began by electroeluting an 865 base pair (bp) Hindlll fragment from pCHIl. We then restricted the pDRIO plasmid with Hindlll. In order to test for homology, we hybridized the pDRIO fragment with nick translated pCHIl fragment as a probe. For a second electrophoresis experiment, we cut pDRIO with EcoRI and Hindlll. We also restricted several different pDRIO plasmids with a variety of deletions. After performing a Southern blot on the second restriction, we were able to determine that the homology for the CHI gene lies in a 1983 bp segment between the multicloning Hindlll site and Pstl site of the insert