Hisbergues Michael, Christiansen Guntram, Rouhiainen Leo, Sivonen Kaarina, Börner Thomas
Department of Biology, Humboldt University, Chausseestrasse 117, 10115 Berlin, Germany.
Arch Microbiol. 2003 Dec;180(6):402-10. doi: 10.1007/s00203-003-0605-9. Epub 2003 Oct 10.
Microcystins are harmful hepatotoxins produced by many, but not all strains of the cyanobacterial genera Anabaena, Microcystis, Anabaena, Planktothrix, and Nostoc. Waterbodies have to be monitored for the mass development of toxic cyanobacteria; however, because of the close genetic relationship of microcystin-producing and non-producing strains within a genus, identification of microcystin-producers by morphological criteria is not possible. The genomes of microcystin-producing cells contain mcy genes coding for the microcystin synthetase complex. Based on the sequence information of mcy genes from Microcystis and Planktothrix, a primer pair for PCR amplification of a mcyA gene fragment was designed. PCR with this primer pair is a powerful means to identify microcystin-producing strains of the genera Anabaena, Microcystis, and Planktothrix. Moreover, subsequent RFLP analysis of the PCR products generated genus-specific fragments and allowed the genus of the toxin producer to be identified. The assay can be used with DNA from field samples.
微囊藻毒素是由蓝藻属的鱼腥藻、微囊藻、颤藻、席藻和念珠藻中的许多菌株(但并非全部)产生的有害肝毒素。必须对水体中有毒蓝藻的大量繁殖进行监测;然而,由于一个属内产生微囊藻毒素的菌株和不产生微囊藻毒素的菌株之间存在密切的遗传关系,因此无法通过形态学标准来鉴定产生微囊藻毒素的菌株。产生微囊藻毒素的细胞基因组包含编码微囊藻毒素合成酶复合体的mcy基因。基于来自微囊藻和席藻的mcy基因序列信息,设计了一对用于PCR扩增mcyA基因片段的引物。用这对引物进行PCR是鉴定鱼腥藻、微囊藻和席藻属中产生微囊藻毒素菌株的有力手段。此外,随后对PCR产物进行的RFLP分析产生了属特异性片段,并能够鉴定毒素产生菌的属。该检测方法可用于野外样品的DNA检测。