Cloning and expression of cDNAs encoding ADP-ribosylation factor in carrot seedling
- 作者：Yuki Asakura ; Eriko Ishigaki ; Ryuji Sugiyama ; Fumiya Kurosaki
- 关键词：GTP-binding protein ; ADP-ribosylation factor ; Gene cloning ; Gene expression ; RT-PCR ; Daucus carota
- 刊名：Plant Science
- 出版年：2007
- 期刊代码：88_01689452
- 类别：abs
- 出版时间：February 2007
- 卷：172
- 期：2
- 页码：189-195
- 文件大小：1066 K
摘要
A homology based strategy yielded two cDNA clones designated arf-001 (1097 bp) and arf-002 (897 bp) from carrot seedlings. These genes contained open reading frames which encode the proteins of 192 and 181 amino acid residues, respectively, with the significant homology to ADP-ribosylation factors of plants, animals and microbial sources. Genomic Southern blot hybridization analyses revealed that both genes showed one main signal, respectively; however, several related genes might be present in carrot genome. The products of these genes obtained by over-expression in Escherichia coli showed the specific binding activity toward GTP. The expression of arf-001 was mainly observed in leaf and stem tissues while arf-002 was appreciably expressed in roots as analyzed by RT-PCR. The transcriptional level of arf-001 showed the transient increase by the exposure of carrot seedlings to ethylene. In contrast, arf-002 appeared to be a house-keeping gene, and its expression level was maintained at constant level upon the treatment with various stimuli and under stress conditions. These results suggest that the arf-001 and 002 play the distinct physiological roles in carrot cells.