VYUŽITIE METÓDY CAPS K DETEKCII POLYMORFIZMU REŠTRIKČNÝCH MIEST GBSSI GÉNU MUTANTNÝCH LÍNIÍ LÁSKAVCA

Abstract

The aim of presented paper was to detect the GBSSI gene. Concretely, the detection of polymorphisms in restriction sites using restriction endonuclease HpaII was performed. Firstly, four parts of the GBSSI gene were amplified in PCR reactions, and then the amplified parts of the GBSSI gene were digested using restriction endonuclease HpaII. The presence of the restriction site 5´CCGG 3´ was confirmed in control genotypes, and in the lines of amaranth exposed to radiation­­–induced mutagenesis as well. The cleavage in all amaranth mutant lines and their controls, two fragments were cleaved. In the part of the GBSSI gene with the size of 1255 bp – 2725bp (primer GBSSI 1) there were two fragments with the size of 1261 bp and 229 bp; in the part of the GBSSI gene with the size of 1 bp – 1098bp (primer GBSSI 3) there were two fragments with the size of 797 bp and 301 bp; and in the part of the GBSSI gene with the size of 2548bp – 3461bp (primer GBSSI 4) there were two fragments with the size of 215 bp and 692 bp.