| Cloning and transcription analysis of Propionyl-CoA Dehydrogenase Gene from Candida rugosa and its expression in Pichia pastoris | |
| Fengli Zhou; Rubing Zhang; Yongguang Zhang; Fan Zhang; Jianhang Zhu. | |
| 2011 | |
| 发表期刊 | Journal of Basic Microbiology
 |
| 期号 | 51页码:1–8 |
| 摘要 | The propionyl-CoA dehydrogenase (PACD) gene was cloned from Candida rugosa by the cDNA RACE technique. The full cDNA of the PACD gene has a length of 1408 bp, which contains a complete open reading frame (ORF) of 1329 bp, coding for 442 amino acids. The cDNA of PACD was cloned into the expression plasmid pPIC9K and transformed into Pichia pastoris GS115. The recombinant protein was purified by Ni-NTA affinity chromatography, and its size was observed to be approximately 49 kDa as estimated by SDS-PAGE. Anti-His antibodies were used to characterise the recombinant PACD by western-blot analysis. The recombinant protein retained the activity of catalysing propionyl-CoA to acryloyl-CoA. The results of dot-blotting hybridisation using a PACD cDNA probe indicated that the PACD mRNA level was modified at different stages: mRNA levels were low for the first 36 h, then increased through 48 h and eventually reached a stable level. These results indicate that propionate induction could significantly activate PACD mRNA expression. Information from this study will be helpful in elucidating the metabolic pathway for 3-hydroxypropionic acid production in C. rugosa. |
| 学科领域 | 生物基化学品 |
| 收录类别 | SCI |
| 语种 | 英语 |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qibebt.ac.cn/handle/337004/1113 |
| 专题 | 生物基材料组群 |
| 推荐引用方式 GB/T 7714 | Fengli Zhou,Rubing Zhang,Yongguang Zhang,et al. Cloning and transcription analysis of Propionyl-CoA Dehydrogenase Gene from Candida rugosa and its expression in Pichia pastoris[J]. Journal of Basic Microbiology,2011(51):1–8. |
| APA | Fengli Zhou,Rubing Zhang,Yongguang Zhang,Fan Zhang,&Jianhang Zhu..(2011).Cloning and transcription analysis of Propionyl-CoA Dehydrogenase Gene from Candida rugosa and its expression in Pichia pastoris.Journal of Basic Microbiology(51),1–8. |
| MLA | Fengli Zhou,et al."Cloning and transcription analysis of Propionyl-CoA Dehydrogenase Gene from Candida rugosa and its expression in Pichia pastoris".Journal of Basic Microbiology .51(2011):1–8. |
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