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微生物学报 2000
EXPRESSION AND ACTIVITY ANALYSIS OF THE HUMAN GLUTATHIONE-S-TRANSFERASE IN LACTOCOCCUS LACTIS
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Abstract:
Theglutathione S transferaseA1cDNAwasamplifiedfromhumanlivertotal RNAsbyRT PCRandwasclonedintoaEscherichiacoliexpressionvectorpET2 3b ,then therecombinantplasmidpET2 3bhgstwasintroducedintoE .coliBL2 1 (DE3)andinduced byIPTG ,thehigh levelexpressionofhGSTA1appearedintheE .colicells.ThecDNAen codinghGSTA1wassubclonedintopMG36e,alactococcalexpressionvector,andintroduced intoLactococcuslactisMG1 36 3byelectroporation .Inthepositivetransformants,thehGS TA1wasexpressedasafusionproteinwhichwasverifiedbySDS PAGEandWesternblot. ThehGSTA1frombothE .coliandL .lactiswaspurifiedbyaffinitychromatographyon glutathione agaroseandallshowedenzymaticactivity .Thepotentialapplicationofthere combinantLactococcuslactisinfunctionalfoodwasalsodisscussed .
