Mazf as a counter-selectable marker for unmarked genetic modification of pichia pastoris
文献类型：期刊论文
| 作者 | Yang, Junjie1,2; Jiang, Weihong1; Yang, Sheng1,3 |
| 刊名 | Fems yeast research
 |
| 出版日期 | 2009-06-01 |
| 卷号 | 9期号:4页码:600-609 |
| 关键词 | Pichia pastoris Mazf Counter-selectable marker Genetic modification |
| ISSN号 | 1567-1356 |
| DOI | 10.1111/j.1567-1364.2009.00503.x |
| 通讯作者 | Yang, sheng(syang@sibs.ac.cn) |
| 英文摘要 | In this study, we demonstrate a novel method for unmarked genetic modification of the methylotrophic yeast pichia pastoris, in which the escherichia coli toxin gene mazf was used as a counter-selectable marker. mazf was placed under the tightly controlled aox1 promoter, and the induced expression of mazf in p. pastoris halted cell growth. a modular plasmid was constructed in which mazf and a zeocin resistance gene acted as counter-selectable and active-selectable markers, respectively, and the mazf-zeor cassette was flanked by two direct repeats for marker recycling. linearized delivery vectors constructed from the modular plasmid were integrated into the p. pastoris genome via homologous recombination, introducing genetic modifications. upon counter-selection with methanol medium, which induces the aox1 promoter, the markers were recycled efficiently via homologous recombination between the direct repeats. we used this method successfully to knock-out the arg1 and met2 genes, knock-in a green fluorescent protein expression cassette, and perform site-directed mutagenesis on the arg1 gene, all without introducing unwanted selection markers. the novel method allows repeated use of the selectable marker gene for multiple modifications and will be a useful tool for p. pastoris studies. |
| WOS关键词 | MESSENGER-RNA INTERFERASE ; SINGLE PROTEIN-PRODUCTION ; PROGRAMMED CELL-DEATH ; ESCHERICHIA-COLI ; BUDDING YEAST ; DISRUPTION ; CLONING ; SYSTEM ; EXPRESSION ; SELECTION |
| WOS研究方向 | Biotechnology & Applied Microbiology ; Microbiology ; Mycology |
| WOS类目 | Biotechnology & Applied Microbiology ; Microbiology ; Mycology |
| 语种 | 英语 |
| WOS记录号 | WOS:000265934800009 |
| 出版者 | WILEY-BLACKWELL PUBLISHING, INC |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2397862 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Yang, Sheng |
| 作者单位 | 1.Chinese Acad Sci, Key Lab Synthet Biol, Inst Plant Physiol & Ecol, Shanghai Inst Biol Sci, Shanghai 200032, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China 3.Chinese Acad Sci, Huzhou Res Ctr Ind Biotechnol, Shanghai Inst Biol Sci, Huzhou, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yang, Junjie,Jiang, Weihong,Yang, Sheng. Mazf as a counter-selectable marker for unmarked genetic modification of pichia pastoris[J]. Fems yeast research,2009,9(4):600-609. |
| APA | Yang, Junjie,Jiang, Weihong,&Yang, Sheng.(2009).Mazf as a counter-selectable marker for unmarked genetic modification of pichia pastoris.Fems yeast research,9(4),600-609. |
| MLA | Yang, Junjie,et al."Mazf as a counter-selectable marker for unmarked genetic modification of pichia pastoris".Fems yeast research 9.4(2009):600-609. |
入库方式： iSwitch采集
来源：中国科学院大学
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。