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作 者:徐芝亮[1,2] 吴灶和[2,3] 简纪常[1,2] 黄郁葱[1,2]
机构地区:[1]广东海洋大学水产学院,广东湛江524088 [2]广东省水产经济动物病原生物学及流行病学重点实验室暨广东高等学校水产经济动物病害控制重点实验室,广东湛江524025 [3]仲恺农业工程学院,广东广州510225
出 处:《水产学报》2012年第9期1450-1456,共7页Journal of Fisheries of China
基 金:广东省科技计划国际合作项目(2009B050700040);国家"九七三"前期专项(2011CB111601);广东省科技计划重大项目(20080100014)
摘 要:运用RAPD分型和BOX分型分子方法研究患病的海水鱼类体内和海水环境的101株哈维氏弧菌的分子遗传多样性。结果显示,从10个RAPD引物中发现只有PM2引物的分辨率和重复性较好,出现15条不同RAPD条带,在相似度65%下,101株哈维氏弧菌可以分为18种不同型;BOX分型出现20条不同的条带,在相似度为40%的情况下,可以把101株哈维式弧菌分15种不同的型;综合RAPD PM2和BOX分型,发现在相似度50%下,101株哈维氏弧菌可分18种型。综合RAPD和BOX分型数据可以很好地克服各自的缺点,得到更加准确的分型结果。Vibrio harveyi is known to be a major pathogen in mariculture animal systems. Disease outbreaks attributed to V. harveyi have been reported in main mariculture animal farm production. In recent years,mass mortalities of many cultured kinds of maricultured fishes, such as Litopenaeus vannamei, Pseudosciaena crocea,and Lutjanus sanguineus have occurred frequently infected by V. harveyi. In this article the 101 isolates of V. harveyi were subjected to random amplified polymorphic DNA (RAPD)-PCR and BOX-PCR analysis to investigate the genetic variability among V. harveyi strains. A total of 10 RAPD primers were designed for their specificity in detecting V. harveyi, and only the primer PM2 was highly reproducible and found suitable to use in RAPD-PCR. The genetic diversity among V. harveyi isolates assessed by RAPD-PCR by PM2 primer yielded 15 different RAPD patterns which clustered the isolates into 18 groups at 65% similarity level. Similarly, BOX-PCR clustered the 20 patterns intol5 groups at 40% similarity. However, with RAPD-PCR and BOX-PCR, 101 V. harveyi could be divided into 18 groups at 50% similarity. We could obtain the accurate results of molecular genetic diversity by both RAPD-PCR and BOX-PCR.
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