Organ-specific gene expression in transgenic potato: the cloning a new promoter of a class I patatin gene Using synthetic oligonucleotide probes homologous to conservative AT-rich motif of patatin genes class I of two differentclones were isolatedfrom a potato genomic library. One of two different genomic clones named X pat 122 was subcloned and analysis 5'-region sequences. Using the chloramphenicol acetyitransferase (CAT) gene as a reporter it has been shown that a L8 kb promoter fragment of the class I patatin gene PAT J 22 provides all the information necessary for both tuber-specific and sucrose-induced expression in leaves in transgenic potato plants.
The bacterial chloramphenicol acety(transferase (CAT) gene was expressed in licorice G. glabra L. suspension protoplasts by electroporation after introduction of the chimeric plasmid pDNt23-CaMV35Snos-cat (pDNt23-root-specific and CaMV 35S promoters, nopaline synthase nos-terminator, and selectable NPT-II gene). Maximum of CAT activity in the licorice protoplasts was observed in SO h after electroporation Together with recent advances in the cell isolation and electroporation procedures, this system allows to study expression of the root-specific promoter introduced into the licorice cells. The level of total flavonoids production in 4 weeks culture was tested in the electroporated cell lines. Additionally, these electroporated cells were treated by elicitors (yeast extract and fungal bioextract). The total flavonoids production increased by 2-5 times in comparison with the non-electroporated licorice cells.
Kanamycin resistant plants of S. tuberosum L. (in vitro-grown) cv. Zarevo were obtained from the cocultivated microtubers with A. tumefaciens. A disarmed binary vector systems containing the neomycin phosphotransferase (NPT II) gene as selectable marker and chloramphenicol acetyltransferase (CAT), as a reporter gene, under control of new patatin promoter class I were utilized. In vitro grown minitubers discs were used as sources of explants to produce transgenic plants on selective medium containing 100 jug/1 kanamycin and CAT enzyme activities were detected.
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