1887

Abstract

A -lactamase-inhibitory protein (BLIP-II) was purified from the culture filtrate of SMF19 and its N-terminal amino acid sequence was determined. A clone containing the gene encoding BLIP-II was selected from a cosmid library by colony hybridization using an oligonucleotide probe based on the N-terminal amino acid sequence of BLIP-II. The gene was isolated and sequenced. Analysis of the nucleotide sequence revealed that the gene consists of 1116 bp and encodes a mature protein of 332 amino acids preceded by a 40 amino acid signal sequence. , expressed under the control of the T7 promoter in , was accumulated in an inactive form in inclusion bodies, but -lactamase-inhibitory activity was recovered after refolding. In addition, was heterologously expressed in TK24 using the /C1 promoter. The BLIP-II protein produced in recombinant strains of was secreted into the culture supernatant in a biologically active form.

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1998-08-01
2024-05-02
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