Abstract
A partial fragment of novel sequence (arr, adriamycin-resistant related) was previously identified using the differential display (DD)-PCR technique with adriamycin-resistant L1210 variant (L1210AdR), which shows a typical multidrug resistant (MDR) phenotypes. The present research shows the isolation of full length arr cDNA sequence. To clone the full length cDNA of arr gene, DD-PCR fragments were subjected to 5'- and 3'-Rapid Amplification of cDNA End (RACE) method. The cloned arr cDNA consisted of 770 bases and contained an open reading frame of 153 bases, encoding a protein of 51 amino acid with the molecular mass of 4 kDa by in vitro translation reactions. Northern blot analysis showed that a 770 bases transcript arr gene was overexpressed in adriamycin-resistant L1210 variant, but not in the parent suggesting that the arr gene may be involved in the adriamycin-resistant phenotypes.
Highlights
The appearance of drug resistant cells is one of the major limiting factor in cancer chemotherapy
These findings suggest that some proteins or genetic alterations are important in the development of antitumor drug resistance in cancer cells
ArrR primers were synthesized on the basis of the original differential display polymerase chain reaction (DD-PCR) fragments
Summary
The appearance of drug resistant cells is one of the major limiting factor in cancer chemotherapy. The differentially expressed cDNA fragments were isolated from L1210AdR cells using DD-PCR as described previously (Kim et al, 1995). To select colonies containing arr-specific DNA fragments amplified by RACE, PCR was carried out using a forward primer (5'-GTTTTCCCAGTCACGACGTTGT-3'), a reverse primer (5'-TGGAATTGTGAGCGGATAAC-3') with vector and an arrR or arrF primer for 5'- or 3'-RACE, respectively.
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