Peculiarities of green fluorescent protein transgene detection in tobacco and maize plants by PCR

Abstract

The aim of the work was to investigate detection of different modifications of the green fluorescent protein gene (gfp) in the transgenic tobacco and maize plants by polymerase chain reaction (PCR). Methods. Total DNA isolation, PCR, electrophoresis of DNA in agarose gel, bioinformatic resources. Results. Three pairs of primers were used for PCR analysis of tobacco and maize containing wild-type gfp or mutant synthetic gene S65Tpgfp. The primer pair gfp1F-gfp1R interacted with the wild-type gfp gene only. The gfp2F-gfp2R primers interacted with the gfp gene of different modifications both in tobacco and maize. The gfp3F-gfp3R primer pair interacted with the modified S65Tpgfp gene in tobacco DNA, but not with maize samples. Conclusions. Primers for detection of heterologous gfp gene, which were both narrowly specific (only one gene modification could be detected), and universal (more than one gene modification could be detected), were verified. It was shown that the primer pair gfp2F-gfp2R was universal for gfp gene detection both in tobacco and maize plants by PCR. The results obtained with gfp2F-gfp2R were reliably reproducible, so this primer pair is recommended for general use.

Keywords

• gfp
• s65tpgfp
• zea maize l.
• nicotiana tabacum l.
• pcr
• transgene detection
• molecular markers.