Luis O. Burzio , Monica Brito , Ana Maria Zarraga , M.A.Q. Siddiqui
{"title":"Assay of chloramphenicol acetyl transferase by high-performance liquid chromatography","authors":"Luis O. Burzio , Monica Brito , Ana Maria Zarraga , M.A.Q. Siddiqui","doi":"10.1016/0735-0651(88)90020-9","DOIUrl":null,"url":null,"abstract":"<div><p>A procedure to measure chloramphenicol acetyl transferase (CAT) activity by reverse-phase high-performance liquid chromotography 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.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"5 1","pages":"Pages 5-8"},"PeriodicalIF":0.0000,"publicationDate":"1988-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(88)90020-9","citationCount":"4","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Gene analysis techniques","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0735065188900209","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 4
Abstract
A procedure to measure chloramphenicol acetyl transferase (CAT) activity by reverse-phase high-performance liquid chromotography 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.
京公网安备 11010802042870号
京ICP备2023020795号-1
分享
求助内容：
应助结果提醒方式：
扫码关注我们
