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水华蓝藻产毒特性的PCR检测法

CHARACTERIZATION OF TOXIC WATERBLOOM-FORMING CYANOBACTERIA BY MODIFIED PCR

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【作者】 潘卉宋立荣刘永定朱运芝沈强

【Author】 PAN Hui, SONG Li-rong, LIU Yong-ding, ZHU Yun-zhi and SHEN Qiang (Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan 430072)

【机构】 中国科学院水生生物研究所!武汉430072

【摘要】 特异引物对(TOX 1P/1F;TOX 2P/2F)用于检测微囊藻毒素合成酶基因mcyB片段在38种水华蓝藻中的分布情况。结果显示,所有能产生微囊藻毒素的微囊藻都有特异扩增条带,非产毒株则没有。几种常规的毒性检测方法验证了PCR方法所获结果的准确性。本研究发展了以全细胞PCR法检测mcyB片断,说明全细胞PCR检测法适用于不同来源的蓝藻材料。结果证明以DNA为基础鉴别产毒和非产毒微囊藻及其他水华蓝藻的方法是可行和实用的。

【Abstract】 The present study surveyed the distribution of microcystin-producing gene- mcyB by PCR among 38 bloom-forming cyanobacterial strains from FACHB-Collection. The results demonstrated that special amplifications were being gotten only from 19 toxic Microcystis strains, whereas it was deficient in nontoxic ones got none. The PCR results showed highly agreement with the toxicity of every strain determined by HPLC, ELISA, and Bioassay respectively. Furthermore, the traditional PCR protocols were simplified by checking the cyanobacteria cells directly instead of the previously used extracted genomic DNA. The whole cells PCR we developed in this study could be successfully applied to cultured algae materials, water samples and lyophilized cyanobacterial cells. Thus, discrimination of toxic and nontoxic Microcystis strains by molecular biological technologies proves to be practicable and efficacious.

【基金】 国家自然科学基金(项目号No.39730380);国家科技部项目(No.DCH-01—001)
  • 【文献出处】 水生生物学报 ,Acta Hydrobiologica Sinica , 编辑部邮箱 ,2001年02期
  • 【分类号】X173
  • 【被引频次】63
  • 【下载频次】614
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