| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
|
|---|
| Outline of Final Research Achievements |
We have constructed the large-deletion mutants of S. avermitilis (SUKA) for expression of exogenous gene clusters for secondary metabolite biosynthesis. The entire biosynthetic gene cluster for polyketide compound is quite large in comparison with biosynthetic gene clusters for other metabolites and some of them are ~ 100 kb. In order to achieve the heterologous expression of these giant biosynthetic gene clusters in SUKA, more efficient methods for gene introduction are indispensable. Most Streptomyces microorganisms strongly restrict DNA prepared in E. coli because they possess unique methyl-specific restriction system. However, S. lividans is not an ideal host since it possesses a weak methyl-specific restriction system. In this study, we have developed efficient heterologous expression system. Procedures are based on integrating BAC vector, a preferable transformation host (S. lividans) and transfer of linear plasmid vector by interspecific mating between S. lividans and SUKA.
|
