Burzio L O, Brito M, Zarraga A M, Siddiqui M A
Instituto de Bioquimica, Facultad de Ciencias, Universidad Austral de Chile, Valdivia.
Gene Anal Tech. 1988 Jan-Feb;5(1):5-8. doi: 10.1016/0735-0651(88)90020-9.
A procedure to measure chloramphenicol acetyl transferase (CAT) activity by reverse-phase high-performance liquid chromatography is described. The antibiotic as well as the acetylated derivatives are well resolved on a Superspher RP-18 column using equal parts of acetonitrile and 10 mM sodium acetate (ph 5.0) as a solvent. Under these conditions, less than 100 pmol of each derivative can be easily detected within 10 minutes, and no radioactive chloramphenicol is needed. The present procedure has been used to measure the activity of the enzyme in extracts of chicken fibroblast transfected with the recombinant plasmid pSV2-cat containing the CAT gene.
本文描述了一种通过反相高效液相色谱法测量氯霉素乙酰转移酶（CAT）活性的方法。在使用等份乙腈和10 mM醋酸钠（pH 5.0）作为溶剂的情况下，抗生素及其乙酰化衍生物在Superspher RP - 18柱上能够得到很好的分离。在这些条件下，10分钟内可以轻松检测到每种衍生物少于100皮摩尔，并且无需使用放射性氯霉素。本方法已用于测量转染了含有CAT基因的重组质粒pSV2 - cat的鸡成纤维细胞提取物中该酶的活性。
