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High-fidelity PCR amplification was performed with Phusion or Q5 DNA polymerase following the manufacturer's instructions (NEB, Ipswich, MA) with a Mobilization of plasmids into S. leeuwenhoekii was performed as established for Streptomyces coelicolor A3(2) by conjugation from E. coli ET12567 (carrying either
26 Oct 2010 We have constructed derivatives of Streptomyces coelicolor M145 as hosts for the heterologous expression of secondary metabolite gene clusters. . Overall, depending on the heterologous gene cluster, M1152 and M1154 produced 20–40 times more antibiotic than the parental strain M145 (Fig. 3).
5 Mar 2014 Structural modification of the GE2270 molecule by genetic engineering may provide a strategy to generate additional compounds of this class for medical use Our laboratory successfully employed Streptomyces coelicolor host strains (M512, M1146, M1152, M1154) [19], for the heterologous expression of
A general approach for the stable amplification of secondary metabolite biosynthetic gene clusters in actinomycetes. •. A panel of powerful Streptomyces coelicolor heterologous hosts for genome mining of natural products. •. A novel DNA editing method for rapidly refactoring gene clusters, including deletion and
in a closely related Streptomyces strain that does not produce chloramphenicol. . S. coelicolor M1152. M145 derivative ?act ?red ?cpk ?cda rpoB(C1298T). 11. S. coelicolor M1581. M1152 containing cosmid pAH91 (S. venezuelae cosmid then treated according to the instructions given in the RNeasy kit (Qiagen,.
15 Sep 2015 The gene cluster cloned from S. roseosporus NRRL 15998 was transferred into several well-developed surrogate hosts including Streptomyces albus J1074, S. coelicolor M1146, S. coelicolor M1152 and S. coelicolor M1154. However, no inhibitory activity against P. aeruginosa was observed with these
16 Sep 2015 A derivative of Streptomyces coelicolor A3(2) designed for the expression of Type III polyketide synthase (PKS) genes was constructed from the .. DNA from E. coli was carried out using standard methods [36] or by using a QIAprep Spin Miniprep Kit (QIAGEN) and following the manufacturer's instructions.
6 Feb 2013 A series of ICZs were recently isolated and identified from a marine-derived actinomycete strain, Streptomyces sanyensis .. Heterologous Expression of the spc Gene Cluster in Streptomyces coelicolor M1152. Heterologous manufacture's instruction and plated on E. coli Top10. The titer of the primary
A) HPLC-UV chromatograms showing production of FK506 in S. coelicolor M1146 after introduction of PAC20N containing the FK506 cluster. Average production levels in exconjugants from strains M1146, M1152 and M1154 were 0.75, 2.81 and 2.06 mg L?1, respectively (Figure 4A), clearly exceeding production in

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February 2018