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Characteristics and gene expression levels of plant transformants modified by one related constructs shearing a gene for proteinase inhibitor.
 BEZUNK, František
Tobacco plants were transformed by bacteria Agrobacterium tumefaciens bearing a plasmid construct with a fusion gene SPI2:GFP and selectable marker NPTII coding for resistance of transformants to antibiotic kanamycin. SPI2:GFP gene was created by a fusion of SPI2- (for serine protease inhibitor) and GFP gene (coding for a green fluorescent protein - GFP) sequences. Transformed tobacco plants were selected on a medium with kanamycin. Positively selected plants were selfed and segregation ratios for NPTII determined for each of genotypes. The presence and function of transgenic DNA was verified by methods of PCR, RT-PCR, Western blot and using the stereomicroscopic fluorescence study of transformants of T0 and T1 generations. This Transgenic genotypes of tobacco Nicotiana tabacum, cv. Petit Havana, SR1 WT showing the expression of the fused gene SPI2:GFP on a level of mRNA and GFP protein were obtained. Thesis was perfomed with a support of the project No. M106030 of Ministry of Education, Youth and Sports of the Czech Republic.
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