[Background] For its fast growing and non-pathogenic property, Mycobacterium smegmatis is used as a model strain for pathogenic Mycobacterium tuberculosis and a workhorse to produce steroid hormones. But it lacks efficient genome deletion system. [Objective] In this study, we established a high efficiency genome deletion system in M. smegmatis assisted by CRISPR-Cas9. [Methods] The Cas9 backbone plasmid pCas9101 was constructed with a tetracycline-inducible codon-optimized cas9 expression operon. Approximat...
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