paraglneSynthetase from Ilscherichia

نویسندگان

  • Akio SuGiyAMA
  • Hiroaki KATo
چکیده

Oyerexpression of the asnA gene from dseherichia coti K-12 coding for asparagine synthetase (EC 6.3.1.1) was achieved with a plasmid, pUNAd37, a derivative of pUC18, in E. cofi. The plasmid was constructed by optimizing a DNA sequence between the promoter and the ribosome binding region. The enzyme, comprising ca. 15% of the totRl soluble protei" in the E. coti cell, was readily purified to apparent homogeneity by DEAE-Cellulofine and Blue-Cel!ulofine column chromatographies. The aminoterminal sequence, amino acid composition, and molecular weight of the purified protein agreed with the predicted yal"es based on the DNA sequence of the gene. Furthermore the natiye moleculftr weight measured by gel filtration ce firmed that asparagine synthetase exists as a dimer of identical subunits.

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تاریخ انتشار 2017