Application of octenidine in inhibiting acetyltransferase and resisting mycobacterium infection
A technology of acetyltransferase and octenidine, applied in the field of pharmacy, can solve problems such as few protease studies
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[0027] 1. Expression and purification of acetyltransferase (Eis) in Mycobacterium tuberculosis
[0028] Expressed according to literature (Chen W, Biswas T, Porter VR, Tsodikov OV, Garneau-Tsodikova S (2011) Unusual regioversatility of acetyltransferase Eis, a cause of drug resistance in XDR-TB.Proc Natl Acad Sci USA 108:9804–9808) and purified acetyltransferase (Eis)
[0029] (1) Transform the strain of Escherichia coli BL21 (Escherichia coli) (DE3) with the pET28a vector containing the Eis gene, and use LB plate medium (containing 50 mg / L kanamycin) to screen for positive clones.
[0030] (2) Pick positive clones on the LB plate culture medium, and transfer to 0.8L LB medium (containing 50mg / L kanamycin) after cultivating overnight at 37°C. When OD600) reached 0.6, 0.1 mM IPTG (Isopropylβ-D-Thiogalactoside) was added and cultured at 16° C. for 16 hours.
[0031] (3) Collect the cells by centrifugation at 5000rpm for 10min and then autoclave the bacteria; centrifuge the sol...
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