Gene sll1454 is important for growth of Synechocystis sp. PCC 6803 on nitrate as the only source of nitrogen

Ronald Stebegg, Georg Schmetterer, Annette Rompel
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Abstract

The gene sll1454 (narB) encodes a nitrate reductase in Synechocystis sp. PCC 6803. Deletion of sll1454 was performed by insertion of a gene coding for a neomycin phosphate transferase (npt) in both directions to the sll1454 ORF resulting in two different mutant strains named sll1454::npt direct and sll1454::npt complementary. Both strains lost the ability to grow on nitrate as the only source of nitrogen, but instead grew at the same rate as the wild type if ammonium was supplied. Trace metal replacement experiments revealed that tungsten ions inhibited the nitrate reductase in Synechocystis sp. PCC 6803, as no growth occurred with nitrate as the sole nitrogen source, if molybdenum ions were replaced equimolarly by tungsten ions. Vanadium ions, on the other hand, did not exert any negative effect under the same conditions and neither did the omission of molybdenum, tungsten or vanadium ions in the growth medium. The co-addition of molybdenum ions to cultures containing tungsten ions restored growth to the same rate as in cultures containing only molybdenum ions or none of them.

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基因 sll1454 对以硝酸盐为唯一氮源的 Synechocystis sp.
sll1454 (narB) 基因编码 Synechocystis sp. PCC 6803 中的硝酸盐还原酶。通过将编码磷酸新霉素转移酶(npt)的基因双向插入 sll1454 ORF 来删除 sll1454,从而产生了两种不同的突变株,分别命名为 sll1454::npt 直接突变株和 sll1454::npt 互补突变株。这两种菌株都失去了以硝酸盐作为唯一氮源的生长能力,但如果提供铵,其生长速度与野生型相同。微量金属置换实验表明,钨离子抑制了 Synechocystis sp. PCC 6803 中的硝酸还原酶,因为如果用钨离子等摩尔置换钼离子,在以硝酸盐为唯一氮源的情况下,菌株不会生长。另一方面,在相同条件下,钒离子没有产生任何负面影响,生长培养基中不含钼离子、钨离子或钒离子也没有产生任何负面影响。在含有钨离子的培养基中同时添加钼离子,可使生长速度恢复到与只含钼离子或不含钼离子的培养基相同的速度。
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