Reith M E, Laudenbach D E, Straus N A
J Bacteriol. 1986 Dec;168(3):1319-24. doi: 10.1128/jb.168.3.1319-1324.1986.
Two mixed oligonucleotide probes derived from conserved regions of the Synechocystis sp. strain PCC 6714 ferredoxin amino acid sequence were utilized to isolate an Anacystis nidulans R2 clone containing the ferredoxin I gene. Nucleotide sequence analysis revealed a 297-base-pair (bp) open reading frame with a deduced amino acid sequence having high homology to other cyanobacterial ferredoxins. Assuming proteolytic cleavage of the initial methionine residue, the molecular weight of the mature A. nidulans R2 ferredoxin was 10,370. The initial methionine residue was preceded by a probable ribosome-binding site sequence, AGGA. Northern hybridization analysis with the cloned ferredoxin gene indicated an RNA transcript of approximately 450 bp. S1 nuclease mapping localized the transcription start site to a position 64 bases upstream from the initial methionine residue. The nucleotide sequence 14 to 8 bp preceding the transcription start site resembled a typical Escherichia coli promoter, but the sequence in the -35 region did not. Southern hybridization detected only a single copy of the ferredoxin sequence in the A. nidulans R2 genome.
从集胞藻属（Synechocystis sp.）菌株PCC 6714铁氧化还原蛋白氨基酸序列保守区衍生而来的两种混合寡核苷酸探针，被用于分离出一个含有铁氧化还原蛋白I基因的巢状集胞藻（Anacystis nidulans）R2克隆。核苷酸序列分析揭示了一个297个碱基对（bp）的开放阅读框，其推导的氨基酸序列与其他蓝细菌铁氧化还原蛋白具有高度同源性。假设起始甲硫氨酸残基发生蛋白水解切割，成熟的巢状集胞藻R2铁氧化还原蛋白的分子量为10370。起始甲硫氨酸残基之前有一个可能的核糖体结合位点序列AGGA。用克隆的铁氧化还原蛋白基因进行的Northern杂交分析表明，有一个约450 bp的RNA转录本。S1核酸酶图谱分析将转录起始位点定位在起始甲硫氨酸残基上游64个碱基处。转录起始位点之前14至8 bp的核苷酸序列类似于典型的大肠杆菌启动子，但-35区域的序列并非如此。Southern杂交在巢状集胞藻R2基因组中仅检测到铁氧化还原蛋白序列的单拷贝。
