%0 Journal Article %A Park, Hyeon Ung %A Lee, Kye Joon %T Cloning and heterologous expression of the gene for BLIP-II, a -lactamase-inhibitory protein from Streptomyces exfoliatus SMF19 %D 1998 %J Microbiology, %V 144 %N 8 %P 2161-2167 %@ 1465-2080 %R https://doi.org/10.1099/00221287-144-8-2161 %K heterologous expression %K -lactamase-inhibitory protein %K cloning %K Streptomyces exfoliatus %I Microbiology Society, %X A -lactamase-inhibitory protein (BLIP-II) was purified from the culture filtrate of Streptomyces exfoliatus SMF19 and its N-terminal amino acid sequence was determined. A clone containing the gene encoding BLIP-II (bliB) 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 bliB 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. bliB, expressed under the control of the T7 promoter in Escherichia coli, was accumulated in an inactive form in inclusion bodies, but -lactamase-inhibitory activity was recovered after refolding. In addition, bliB was heterologously expressed in Streptomyces lividans TK24 using the me/C1 promoter. The BLIP-II protein produced in recombinant strains of S. lividans was secreted into the culture supernatant in a biologically active form. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-144-8-2161