Active Ingredients: Ciprofloxacin
Natural Transformation Natural transformation biphasic method was performed following standard procedure as described by Wang and Taylor 1990. Cloning, expression, and purification of rCj 0843 c were performed using the procedures described in our previous publication Lin et al.
The plasmid pET 28 b-Cj 0843 c in the E. Approximately 4 mg of highly purified rCj 0843 c was used for the production of rabbit polyclonal antisera by Pacific Immunology Corp Ramona, CA. Pre- and post-immune serum samples were analyzed by immunoblotting against pure rCj 0843 c to confirm the generation of specific Cj 0843 c antibody.
Enzymatic Activity of rCj 0843 c The LT activity of rCj 0843 c was evaluated using turbidimetric assay as reported before with minor modification Caldentey and Bamford, 1992.
In brief, the isogenic Cj 0843 c mutant of C. Then 3 ml of chloroform was added in the suspension, which was kept at room temperature for 30 min with gently shaking.
Subsequently, the chloroform-extracted cells the crude cell walls were pelleted by centrifugation at 13,000 rpm for 5 min, washed once with 50 mM Tris-HCl buffer pH 7.
To perform turbidimetric assay, the crude cell walls were adjusted to final OD 450 nm of approximately 0. The decrease in turbidity at 450 nm was determined after 2.
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