用于确定鱼腥藻属PCC 7120菌株异形胞和营养细胞中基因差异表达的载体。
Vectors for determining the differential expression of genes in heterocysts and vegetative cells of Anabaena sp. strain PCC 7120.
作者信息
Bauer C C, Haselkorn R
机构信息
Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637, USA.
出版信息
J Bacteriol. 1995 Jun;177(11):3332-6. doi: 10.1128/jb.177.11.3332-3336.1995.
Abstract
Plasmid vectors were constructed to study promoters of the cyanobacterium Anabaena sp. strain PCC 7120. Plasmid pCCBSelect contains the promoterless reporter genes in the order cat-nifHDK. In pCCBSelect/a, the nifHDK operon precedes the cat gene. Putative promoter sequences were cloned into a polylinker region upstream of the reporter genes. Activity in heterocysts was determined by complementation of a strain containing a deletion of the nifH gene. Activity in vegetative cells was determined by measuring resistance to chloramphenicol. The promoter of the nifHDK operon was active only in heterocysts; the promoter of the nifJ gene was active only in iron-depleted medium; and the promoters of the psbB gene, the ntcA gene, and a newly found transcription factor gene were all active in both cell types.
摘要
构建了质粒载体以研究蓝藻鱼腥藻PCC 7120菌株的启动子。质粒pCCBSelect含有无启动子的报告基因，顺序为cat - nifHDK。在pCCBSelect/a中，nifHDK操纵子位于cat基因之前。将推定的启动子序列克隆到报告基因上游的多克隆位点区域。通过对含有nifH基因缺失的菌株进行互补来确定异形胞中的活性。通过测量对氯霉素的抗性来确定营养细胞中的活性。nifHDK操纵子的启动子仅在异形胞中具有活性；nifJ基因的启动子仅在缺铁培养基中具有活性；psbB基因、ntcA基因和一个新发现的转录因子基因的启动子在两种细胞类型中均具有活性。
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