Premium
Retention of antigenicity by a fragment of Aeromonas salmonicida 70‐kDa serine protease which includes the primary substrate binding site expressed as β‐galactosidase hybrid proteins
Author(s) -
BENNETT A. J.,
WHITBY P. W.,
COLEMAN G.
Publication year - 1992
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.1992.tb00679.x
Subject(s) - aeromonas salmonicida , serine protease , biology , tmprss6 , microbiology and biotechnology , antigenicity , masp1 , serine , protease , biochemistry , fusion protein , protein subunit , enzyme , gene , antibody , recombinant dna , bacteria , genetics
. A 587 bp Pvu II restriction fragment from the 70‐kDa Aeromonas salmonicida serine protease gene, containing the‘active serine’ site sequence of the enzyme, has been cloned into the Sma I restriction site of pUEX2 which on expression, in Escherichia coli DH5α, produced a 142‐kDa hybrid protein in high yield. The hybrid consisted of a fusion between an essentially complete β‐galactosidase subunit and approximately one‐third of the serine protease. A further 42‐kDa hybrid was constructed from the same fragment of serine protease fused to a truncated α‐galactosidase subunit. Both fusion proteins were shown to possess recognizable epitopes by dot blotting against rabbit anti‐ A. salmonicida 70‐kDa serine protease antibody.