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Detection and Expression of Biosynthetic Genes in Actinobacteria ...

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_____________________________________________________________________Table 35: Cerylid act<strong>in</strong>obacterial isolates A1488 <strong>and</strong> A3023 nucleotide <strong>and</strong>translated am<strong>in</strong>o acid sequence, percentage similarity with aromaticPKS genes us<strong>in</strong>g gapped-FASTA database search ________________ 80Table 36: Secondary metabolite biosynthetic gene nucleic acid <strong>and</strong> am<strong>in</strong>o acidsequences <strong>in</strong>corporated <strong>in</strong>to the design <strong>of</strong> the ole01f <strong>and</strong> ole01r PCRprimers _________________________________________________ 82Table 37: Relative am<strong>in</strong>o acid positions <strong>and</strong> codon degeneracies <strong>of</strong> ole01f <strong>and</strong>ole01r primers with<strong>in</strong> the sequence ___________________________ 83Table 38: Relative nucleic acid positions <strong>and</strong> <strong>of</strong> ole01f <strong>and</strong> ole01r primers <strong>and</strong>variable nucleotides with<strong>in</strong> the sequence _______________________ 83Table 39: Comparison <strong>of</strong> PCR amplified KS gene from Cerylid cultures A0350<strong>and</strong> A1113 sequence percentage similarity with mPKS genes us<strong>in</strong>g theFASTA database search ____________________________________ 85Table 40: Secondary metabolite biosynthetic gene nucleic acid <strong>and</strong> am<strong>in</strong>o acidsequences <strong>in</strong>corporated <strong>in</strong>to the design <strong>of</strong> the strD01f <strong>and</strong> strD01r PCRprimers _________________________________________________ 88Table 41: Relative am<strong>in</strong>o acid positions <strong>and</strong> codon degeneracies <strong>of</strong> strD01f<strong>and</strong> strD01r primers with<strong>in</strong> the sequence _______________________ 89Table 42: Relative nucleic acid positions <strong>and</strong> <strong>of</strong> strD01f <strong>and</strong> strD01r primers<strong>and</strong> variable nucleotides with<strong>in</strong> the sequence____________________ 89Table 43: Comparison <strong>of</strong> nucleotide <strong>and</strong> am<strong>in</strong>o acids from amplifieddTDP-glucose synthase gene from Streptomyces grtiseus DSM40236Type Stra<strong>in</strong> with dTDP-glucose synthase from different act<strong>in</strong>obacterialstra<strong>in</strong>s ___________________________________________________ 91Table 44: Isopenicill<strong>in</strong> N synthase genes with their correspond<strong>in</strong>g nucleicacid <strong>and</strong> am<strong>in</strong>o acid sequences <strong>in</strong>corporated <strong>in</strong>to the design <strong>of</strong> thepcb03f <strong>and</strong> pcb03r PCR primers______________________________ 92Table 45: Relative am<strong>in</strong>o acid positions <strong>and</strong> codon degeneracies <strong>of</strong> pcb03f<strong>and</strong> pcb03r primers with<strong>in</strong> the sequence ________________________ 93Table 46: Relative nucleic acid positions <strong>and</strong> <strong>of</strong> pcb03f <strong>and</strong> pcb03r primers<strong>and</strong> variable nucleotides with<strong>in</strong> the sequence ____________________ 94Table 47: Comparison <strong>of</strong> nucleotide <strong>and</strong> am<strong>in</strong>o acid sequences fromisopenicill<strong>in</strong> N synthase (pcbC) gene with sequenced Streptomycesgriseus NRRL3851 <strong>and</strong> Streptomyces cattleya ATCC8507Type Stra<strong>in</strong>s ______________________________________________ 95Table 48: Summary <strong>of</strong> PCR screen<strong>in</strong>g on Cerylid environmental isolates ______ 96Table 49: Antibacterial <strong>and</strong> antifungal activities <strong>of</strong> extracts from the Cerylidenvironmental act<strong>in</strong>obacterial cultures: plug <strong>and</strong> well type diffusionassays ___________________________________________________ 97Table 50: Fermentation analysis, depict<strong>in</strong>g the antimicrobial activities forsupernatant <strong>and</strong> mycelial extracts <strong>of</strong> the Cerylid environmentalact<strong>in</strong>obacterial isolates _____________________________________ 99Table 51: Comparison <strong>of</strong> fermentation conditions for bioactive metabolitescreen<strong>in</strong>g ________________________________________________ 100Table 52: Characterization <strong>of</strong> ethyl acetate extracted metabolites from A2707culture on different solid-substrate fermentation media ____________ 101Table 53: Well-Type anti-bacterial assay <strong>of</strong> mycelium extracted metabolitesfrom act<strong>in</strong>obacterial liquid-oil fermentations after 10 days <strong>in</strong>cubation_ 102_____________________________________________________________________viii

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