Cloning and Analysis of the Auxin Efflux Carrier Gene in Bitter Gourd
| Autor: | Shin-Han Lee, 李思涵 |
|---|---|
| Rok vydání: | 2002 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 90 The putative auxin efflux carrier gene MCm-AEC1 from bitter gourd was isolated from a genomic library constructed in the λEMBL3 vector by screening with pMAEC28 and pMAEC93 cDNAs. Approximately 1.5x106 plaque-forming units were screened by plaque hybridization and 34 putative clones were purified individually. According to the results of dot hybridization and restriction endonuclease site map, the auxin efflux carrier genomic clones were classified into two groups. One of the genomic clones corresponding to pMAEC28,λMCm-AEC1-34, was completely sequenced and characterized. MCm-AEC1 gene in λMCm-AEC1-34, spanning 3,206 base pairs, contains six exons and five introns with consensus AG-GT dinucleotides locating at their boundaries. The amino acid sequences of bitter gourd auxin efflux carrier shows 45.6~76.61% homology to auxin efflux carrier of Arabidopsis, Brassica and rice. The first 26 amino acids are predicted to be a putative signal peptide. Four conserved repeats exist in the polypeptide. The putative TATA and CAAT boxes are 449~455 bp and 570~574 bp upstream from the translation start site in MCm-AEC1, respectively. Several conserved elements responsive to auxin, ethylene, light, salicylic acid and wounding are found in the promoter region. Northern blot analysis indicated that the expression of bitter gourd auxin efflux carrier gene MCm-AEC1 was induced by 10-3 Ｍ IAA, IBA, NAA and 2,4-D. No obvious gene expression was detected in other treatments. To analyze the promoter activity, 3 kb promoter fragment was fused to GUS reporter gene coding sequence for further studies. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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