Chapter title |
Genetic Techniques for Manipulation of the Phytosterol Biotransformation Strain Mycobacterium neoaurum NRRL B-3805
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Chapter number | 7 |
Book title |
Methods in Molecular Biology
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Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7183-1_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7182-4, 978-1-4939-7183-1
|
Authors |
Loraine, Jessica K., Smith, Margaret C. M., Jessica K. Loraine, Margaret C. M. Smith |
Abstract |
Mycobacterium neoaurum is a saprophytic, soil-dwelling bacterium. The strain NRRL B-3805 converts phytosterols to androst-4-ene-3,17-dione (androstenedione; AD), a precursor of multiple C19 steroids of importance to industry. NRRL B-3805 itself is able to convert AD to other steroid products, including testosterone (Ts) and androst-1,4-diene-3,17-dione (androstadienedione; ADD). However to improve this strain for industrial use, genetic modification is a priority. In this chapter, we describe a range of genetic techniques that can be used for M. neoaurum NRRL B-3805. Methods for transformation, expression, and gene knockouts are presented as well as plasmid maintenance and stability. |
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