一株溶血性蜡状芽孢杆菌wh2015全基因组测序分析及其溶没血基因筛选-水产养殖专业毕业论文.docx

Genome Genome Analysis of a Hemolytic Bacillus cereus and Screen its Hemolysis Genes Abstract Bacillus cereus is a Gram—positive and spore—forming bacterium which can be found in foods，rice dishes，eggs，spices，meat，dairy products and even the drugs．Severe food poisoning accidents have been reported to be caused by the B．cereus with vomiting and diarrhea in many countries，including the USA，Canada and the Netherlands． In this study,a hemolytic bacterium strain isolated from a farmed turbot，and now Scophthalmus maximus(turbot)became one of the dominant species in mariculture fishes in China with an annual production more than 60000 tons，during the rapid development of turbot aquaculture，fish diseases emerged and became serious problems，many pathogenic bacteria were reported to cause hemolysis of the host，the isolates could be a consistent pathogenic germs，SO we have a following study on the hemolytic bacteria： 1．Based on morphological and phylogenetic analyses，the isolate was identified as Bacillus cereus and was named as WH20 1 5 according to the isolation place and time． 2．Blood agar hemolytic assay confirmed that the WH20 1 5 strain could lyse the sheep red blood cells．electronic scanning electron microscopy(sem)analysis confirmed B． cereus WH2015 have a certain ability to damage the blood cells． 3．We conducted genome sequencing and annotation of WH20 1 5 strain．The results showed that B．cereus WH20 1 5 has a genome size around 5．8 Mb，with an average G+C content of 35％，6 568 annotated protein—coding genes and 98 tRNA sequence．Comparing with other sequenced B．cereus strains，the WH20 1 5 genome possesses almost all of the core hemolytic genes，a total of 23 virulence factor genes，including 1 HBL operon gene，a hlylII gene，strongly suggest that its pathogenesis mechanisms are similar to other reported B．cereus isolates． 4．To test and verify the function of hemolytic genes，this paper also successfully construct prokaryotic expression plasmid pET-28a—hblC，pET-28a—hblD，pET-