摘要
CytoehromeP450norgenewasclonedintotheexpressionvectorpET-28toyieldtherecombinantexpressionplasmidpET-P450nor,whichcoulddirectthesynthesisofaeukaryoticderivedproteininEscherichiacoliBL21.Thevectorallowsoverproductionandsingle-steppurificationof(His)6-taggedcytoehromeP450norbythefacifitationofmetal(Ni^2+)chelateafl]nitychromatography.TheexpressionlevelofcytoehromeP450norwashighat30℃afterIPTGinduction.SDSPAGEandWesternblotanalysisshowedaspecificband(about43kDa).TheoverproducedcytochromeP450norwaspurifiedtoeleetrophoretichomogeneitywithin2.5handabout20.8mgpurifiedproteinwasobtainedfrom2Lcellculture.TheproliferationofSSMC-7721celllinecouldbeinhibitedbycytoehromeP450nor.Rabbitpolyclonalantibodies(titerover64000)wereproducedagainstrecombinantcytoehromeP450norandprovedtobeveryusefulforimmunoblottingstudy.AvailabilityofthisexpressionsystemandspecificantibodiesshouldfacilitatecharacterizationoftheroleofcytochromeP450norinthemetabolismofNO.
出版日期
2004年02月12日（中国期刊网平台首次上网日期，不代表论文的发表时间）
