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1

Differential display - the method of searching for specifically expressed genes

100%
Dabrowska G.
Biotechnologia
|
2001
|
issue 3
124-133
EN
Identification of differentially expressed genes is one of the major challenges in molecular biology. Several techniques allow the cloning of such sequences. However, methods such as differential hybridization are time-consuming and require large amounts of mRNA. Recently a new approach has been successfully developed: differential display by polymerase chain reaction (DD PCR). This technique has been proven to be highly effective in identifying sequences that are differentially expressed in various cell types.
2

Screening of the EST database for identification of the complete cDNA sequence of the gene encoding aquaporin of Pharbitis nil Choisy (PnPIP1)

63%
Dabrowska G., Mordaka P. M.
Biotechnologia
|
2008
|
issue 2
190-198
EN
Aquaporins are membrane proteins that facilitate water transport across the membranes in various microorganisms, plants and animals. Plant aquaporins are divided into four groups based on the amino acid sequence similarities and intracellular localization: plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin-like intrinsic proteins (NIPs) and small basic intrinsic proteins (SIPs). We found 35 EST sequences homologous to 3' and 5' termini of the partial cDNA of P. nil in the GenBank NCBI database. cDNA encoding full length aquaporin of P. nil was cloned with the use of the reverse transcription-polymerase chain reaction (RT-PCR). The 1189 bp full length cDNA sequence of aquaporin P. nil (PnPIP1) was obtained. Analysis of protein hydropathy indicated that cloned part of PnPIP1 contained the NPA motif (Asn-Pro-Ala) that is present in all known aquaporins. The amino acid sequence of the PnPIP1 protein exhibits 90, 89 and 88% sequence similarity to Petunia x hybrida, Nicotiana excelior and Fraxinus excelsior aquaporins respectively. We showed that the EST database is a useful tool for identification of the complete cDNA of known genes.
3

Application of callus tissue in the studies on molecular mechanism of flower induction

51%
Szyp I., Dabrowska G., Tretyn A.
Biotechnologia
|
2001
|
issue 2
165-169
EN
Flowering is a crucial turning point in the life cycle of most plants. The process of flowering is controlled by external factors such as light and temperature. Floral induction is the first step in the transition from the vegetative to reproductive stage of development. In the photoperiodically sensitive plants this process is regulated by the duration of light and darkness during a 24-h cycle. The aim of our study was to determine whether the undifferentiated callus tissue obtained from cotyledons, is suitable for molecular investigations on the mechanisms of flower induction. The callus tissue was obtained from cotyledons of Pharbitis nil plants, which were cultivated in inductive or non-inductive conditions. The callus obtained after two subcultures was used for isolation of RNA. The total RNA was extracted as described by Chomczynski (1993). We have examined the changes in the pattern of RNA in these two types of callus, using the technique of differential display by the polymerase chain reaction (PCR). Differential display is a method for the identification and cloning of differentially expressed eucaryotic genes. We have found the differences between patterns of RNA derived from callus tissue cultivated under non-inductive conditions and callus tissue cultivated under inductive conditions. In conclusion we can suggest that the tested callus preserved the information on the photoperiodic induction in cells. The process of undifferentiation did not result in the loss of the properties acquired by cotyledon tissue during the photoperiodic treatment.
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