Chung Chih-Ching, Hwang Sheng-Ping L, Chang Jeng
Institute of Marine Biology, National Taiwan Ocean University, Keelung 20224, Taiwan, Republic of China.
Appl Environ Microbiol. 2005 Dec;71(12):8744-51. doi: 10.1128/AEM.71.12.8744-8751.2005.
A novel death-specific gene, ScDSP, was obtained from a death stage subtraction cDNA library of the diatom Skeletonema costatum. The full length of ScDSP cDNA was 921 bp in length, containing a 699-bp open reading frame encoding 232 amino acids and two stretches of 66 and 156 bp in the 5' and 3' untranslated regions, respectively. Analysis of the peptide structure revealed that ScDSP contained a signal peptide domain, a transmembrane domain, and a pair of EF-hand motifs. When S. costatum grew exponentially at a rate of 1.3 day(-1), the ScDSP mRNA level was at 2 mumol . mole 18S rRNA(-1). In contrast, when the culture entered the death phase with a growth rate decreasing to 0.5 day(-1), ScDSP mRNA increased dramatically to 668 mumol . mole 18S rRNA(-1), and a high degree of DNA fragmentation was simultaneously observed. Under the influence of a light-dark cycle, ScDSP expression in both exponential and stationary phases clearly showed a diel rhythm, but the daily mean mRNA level was significantly higher in the stationary phase. Our results suggest that ScDSP may play a role in the molecular mechanism of self-destructive autolysis in phytoplankton under stress.
从硅藻中肋骨条藻死亡阶段消减cDNA文库中获得了一个新的死亡特异性基因ScDSP。ScDSP cDNA全长921 bp,包含一个699 bp的开放阅读框,编码232个氨基酸,5'和3'非翻译区分别有两段长度为66 bp和156 bp的序列。对该肽结构的分析表明,ScDSP包含一个信号肽结构域、一个跨膜结构域和一对EF-手基序。当中肋骨条藻以1.3 d⁻¹的速率指数生长时,ScDSP mRNA水平为2 μmol·mol⁻¹ 18S rRNA⁻¹。相比之下,当培养物进入死亡阶段,生长速率降至0.5 d⁻¹时,ScDSP mRNA急剧增加至668 μmol·mol⁻¹ 18S rRNA⁻¹,同时观察到高度的DNA片段化。在光暗循环的影响下,ScDSP在指数期和稳定期的表达均明显呈现出昼夜节律,但稳定期的日均mRNA水平显著更高。我们的结果表明,ScDSP可能在应激条件下浮游植物自毁性自溶的分子机制中发挥作用。