Detection and Expression of Biosynthetic Genes in Actinobacteria ...

BERVANAKIS, G.Chapter 1: INTRODUCTIONTable 9: PCR Screening for SMBG in Actinobacteria spp.Actinobacteria Biosynthetic Source ReferenceGenesS.coelicolor A3(2) Type I PKS a gDNA library Kuczek et al., 1997S. caelestis Type I PKS gDNA library Kakavas et al., 1997S. antibioticus Type I PKS gDNA library Shah et al., 2000S.hygroscopicus.var Type I PKS gDNA library Wu et al., 2000ascomyceticusS. venezuelae Type I PKS gDNA library Xue et al., 2000Streptomyces spp. Type II PKS Soil Metsa-Ketela et al,1999Streptomyces spp. Type II PKS Soil Seow et al., 1997Streptomyces spp. Type II PKS Soil Morris et al., 1999Streptomyces spp. Type II PKS cDNA Philaniappan et al.,1999Streptomyces spp. Type II PKS cDNA Ziermann andBetlach, 1999Actinomycete spp. NGDH* gDNA library Decker et al., 1996S.antibioticus Tü99 NGDH gDNA library Jin-Cheol et al., 1999S. globisporus C- NGDH gDNA library Liu and Shen, 20001027Actinobacteria spp. NGS~ cDNA Hyun et al., 2000Streptomyces spp. pcbC ^ cDNA Krallis and Kirby,1998S. sulfonofaciens pcbC cDNA Niebla et al., 1999Amycolatopsis Glycosyltransferase gDNA library Pelzer et al., 1999mediterraneiS.venezuelaeISP5230Halogenase gDNA Piraee et al., 2002Actinobacteria spp. Aminotransferase Soil Nagaya et al., 2005aPKS = indicates one or more polyketide synthase genes* NGDH = dNTP-glucose 4,6 dehydratase involved in deoxysugar biosynthesis~ NGS = dNTP-glucose synthase involved in deoxysugar biosynthesis^ pcbC = isopenicillin N synthase gene involved in β-lactam biosynthesis1.6.4 Accessing Secondary Metabolite Diversity from Uncultured SoilMicroorgansimsThe culturability of a number of terrestrial microorganisms present in soil habitats hasproven difficult due to a lack of understanding of the physiology and the inability ofcurrent techniques to culture these microorganisms (Amann et al., 1995; Rondon etal., 2000). Accessing the metabolic diversity without culturing microorganisms can beachieved by using a shotgun cloning approach (Figure 15) which involves isolating_____________________________________________________________________35