Campbell W S, Laudenbach D E
Department of Plant Sciences, University of Western Ontario, London, Canada.
J Bacteriol. 1995 Feb;177(4):964-72. doi: 10.1128/jb.177.4.964-972.1995.
By using an oligonucleotide probe constructed from a conserved region of amino acids located in the carboxyl-terminal end of superoxide dismutase (SOD) proteins, four SOD genes were cloned from the cyanobacterium Plectonema boryanum UTEX 485. One of these genes, designated sodB, encoded an FeSOD enzyme, while the remaining three genes, designated sodA1, sodA2, and sodA3, encoded MnSOD enzymes. To investigate the expression of these four genes, total cellular RNA was isolated from P. boryanum UTEX 485 cells grown under various conditions and RNA gel blot analysis was carried out. Results indicated that sodB and sodA1 were constitutively expressed, although sodB expression was partially repressed in cells grown under conditions of iron stress. sodA2 transcripts, which were not detectable in control cells, accumulated to high levels in cells treated with methyl viologen or in cells grown under conditions of iron or nitrogen stress. However, under microaerobic conditions, iron and nitrogen stress failed to induce sodA2, indicating that multiple factors affect the regulation of sodA2. While discrete transcripts were not detected for sodA3, hybridization was observed under a number of conditions, including those which increased the accumulation of sodA2 transcripts. Additionally, there were high levels of the sodA3 transcript detected in a P. boryanum UTEX 485 mutant strain resistant to methyl viologen treatment.
通过使用由超氧化物歧化酶(SOD)蛋白羧基末端保守氨基酸区域构建的寡核苷酸探针,从蓝藻颤藻Plectonema boryanum UTEX 485中克隆出四个SOD基因。其中一个基因命名为sodB,编码一种铁超氧化物歧化酶(FeSOD),而其余三个基因命名为sodA1、sodA2和sodA3,编码锰超氧化物歧化酶(MnSOD)。为了研究这四个基因的表达情况,从在各种条件下生长的颤藻Plectonema boryanum UTEX 485细胞中分离出总细胞RNA,并进行RNA凝胶印迹分析。结果表明,sodB和sodA1组成型表达,尽管在铁胁迫条件下生长的细胞中sodB表达受到部分抑制。在对照细胞中未检测到的sodA2转录本,在用甲基紫精处理的细胞或在铁或氮胁迫条件下生长的细胞中积累到高水平。然而,在微需氧条件下,铁和氮胁迫未能诱导sodA2,表明多种因素影响sodA2的调控。虽然未检测到sodA3的离散转录本,但在许多条件下都观察到杂交信号,包括那些增加sodA2转录本积累的条件。此外,在对甲基紫精处理具有抗性的颤藻Plectonema boryanum UTEX 485突变株中检测到高水平的sodA3转录本。