Method for increasing yield of erythromycin through saccharopolyspora erythraea SACE_1906 gene pathway
A technology of Saccharopolyspora erythromycetes and erythromycin, which is applied in the field of genetic engineering and can solve the problem of the absence of regulatory genes in synthetic gene clusters
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[0031] 1 Materials and methods:
[0032] 1.1 Strains, plasmids and growth conditions
[0033] The bacterial strains and plasmids used in the examples are shown in Table 1. Escherichia coli was cultured in liquid LB medium at 37°C or LB solid plate with no resistance or corresponding resistance added with 1.25% agar. The erythromycin-producing bacterium Erythropolyspora saccharopolyspora and its engineered strains were cultured on tryptone soybean broth (TSB) medium at 30°C or R3M solid plates containing 2.2% agar without resistance or corresponding resistance.
[0034] 1.2 Materials, DNA manipulation and sequencing
[0035] PEG3350, lysozyme, TES, casamino acids, thiostrepton, and apramycin were purchased from Sigma. TSB, yeast extract, and peptone were purchased from Oxoid. Glycine, agar powder, sodium chloride, and other biological reagents were purchased from reagent companies. The general operating techniques for Escherichia coli and Saccharopolyspora red mold follow ...
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