期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 59, 期 2, 页码 181-188出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2004.06.009
关键词
filamentous cyanobacteria; GFP; transcriptional reporter vectors; akinetes; hormogonia; heterocysts; pSUN 119; pSUN202
资金
- NIGMS NIH HHS [GM63787, S06 GM048680, GM48680] Funding Source: Medline
Two transcriptional reporter shuttle vectors were constructed for the filamentous cyanobacterium Nostoc punctiforme using the green fluorescence protein (GFP) reporter. Both the ampicillin-and kanamycin-resistant versions of the plasmid allow promoters to be directionally cloned into a multiple cloning site preceding a promoterless gfp gene using an Escherichia coli host. The ability of the self-replicating shuttle plasmids to report cell-type-specific gene expression in N. punctiforme was tested by cloning promoters expressed in normal vegetative cells, nitrogen-fixing heterocysts and spore-like akinetes. A P-psaC reporter gene fusion was expressed in vegetative cells and not in heterocysts, whereas GFP driven from P-hetR was found highly expressed in heterocysts. GFP expression driven by the promoter for the N. punctiforme homologue of the akinete-specific gene avaK was expressed in developing akinetes. Decreased expression of GFP from the P-psaC reporter in hormogonia was also observed. The results demonstrate the utility of these GFP vectors to study cell-type-specifie gene expression in differentiating filamentous cyanobacteria. (C) 2004 Elsevier B.V. All rights reserved.
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