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首页> 外文期刊>Harmful Algae >Validation of the detection of Alexandrium species using specific RNA probes tested in a microarray format: Calibration of signal using variability of RNA content with environmental conditions
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Validation of the detection of Alexandrium species using specific RNA probes tested in a microarray format: Calibration of signal using variability of RNA content with environmental conditions

机译：使用以微阵列格式测试的特定RNA探针验证亚历山大菌种的检测：使用RNA含量随环境条件的变化对信号进行校准

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摘要

The dinoflagellate genus Alexandrium contains several toxin producing species and strains, which can cause major economic losses to the shell fish industry. It is therefore important to be able to detect these toxin producers and also distinguish toxic strains from some of the morphologically identical non-toxic strains. To facilitate this DNA probes to be used in a microarray format were designed in silico or developed from existing published probes. These probes targeted either the 18S or 28S ribosomal ribonucleic acid (rRNA) gene in Alexandrium tamarense Group Ⅰ, Group Ⅲ and Group Ⅳ, Alexandrium ostenfeldii and Alexandrium minutum. Three strains of A. tamarense Group Ⅰ, A. tamarense Group Ⅲ, A. minutum and two strains of A. ostenfeldii were grown at optimal conditions and transferred into new environmental conditions changing either the light intensity, salinity, temperature or nutrient concentrations, to check if any of these environmental conditions induced changes in the cellular ribonucleic acid (RNA) concentration or growth rate. The aim of this experiment was the calibration of several species-specific probes for the quantification of the toxic Alexandrium strains. Growth rates were highly variable but only elevated or lowered salinity significantly lowered growth rate for A. tamarense Group Ⅰ and Group Ⅲ; differences in RNA content were not significant for the majority of the treatments. Only light intensity seemed to affect significantly the RNA content in A. tamarense Group Ⅰ and Group Ⅲ, but this was still within the same range as for the other treatments meaning that a back calibration from RNA to cell numbers was possible. The designed probes allow the production of quantitative information for Alexandrium species for the microarray chip.
机译：鞭毛藻亚纲属包含几种产毒素的物种和菌株，可能对贝类养殖业造成重大经济损失。因此，重要的是能够检测出这些毒素产生者，并将有毒菌株与一些形态相同的无毒菌株区分开。为了促进该DNA探针以计算机芯片设计或由现有公开的探针开发，以微阵列形式使用。这些探针分别针对塔玛亚历山大藻第一，第三和第四组，奥斯滕费尔德亚历山大和小亚历山大藻的18S或28S核糖体核糖核酸（rRNA）基因。在最佳条件下培养了三株塔玛生菌A. tamarenseⅠ，塔玛生菌A. tamarenseⅢ，最小拟南芥A. minutum和两株奥斯汀A. ostenfeldii，并将其转移到新的环境条件中，从而改变了光强度，盐度，温度或营养浓度，检查是否有任何这些环境条件引起了细胞核糖核酸（RNA）浓度或生长速率的变化。本实验的目的是对几种物种特异性探针进行校准，以定量毒性亚历山大菌株。生长速度变化很大，但只有盐度的升高或降低才显着降低了塔玛农AⅠ和Ⅲ组的生长率。在大多数治疗中，RNA含量的差异并不显着。 tamarenseⅠ组和Ⅲ组中只有光强度似乎会显着影响RNA含量，但这仍与其他处理方法处于同一范围内，这意味着从RNA到细胞数的反向校准是可能的。设计的探针允许产生用于微阵列芯片的亚历山大属物种的定量信息。

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  • 来源
    《Harmful Algae》 |2014年第7期|17-27|共11页
  • 作者

    Joe D. Taylor; Jessica U. Kegel; Jane M. Lewis; Linda K. Medlin;

  • 作者单位

    Faculty of Science and Technology, University of Westminster, 115 New Cavendish Street, London W1W 6UW, UK,Marine Biological Association of the UK, The Laboratory, The Citadel, Plymouth PL1 2PB, UK;

    Marine Biological Association of the UK, The Laboratory, The Citadel, Plymouth PL1 2PB, UK;

    Faculty of Science and Technology, University of Westminster, 115 New Cavendish Street, London W1W 6UW, UK;

    Marine Biological Association of the UK, The Laboratory, The Citadel, Plymouth PL1 2PB, UK;

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  • 正文语种 eng
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  • 关键词

    Microarray; Phylochip; rRNA probes; Toxic algae; Alexandrium; MIDTAL; RNA content;

    机译：微阵列;Phylochip;rRNA探针;有毒藻类;亚历山大;MIDTAL;RNA含量;

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