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Cloning and sequencing of a Clostridium perfringens sialidase gene
Author(s) -
Roggentin Peter,
Rothe Bernd,
Lottspeich Friedrich,
Schauer Roland
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80219-9
Subject(s) - clostridium perfringens , cloning (programming) , sialidase , chemistry , gene , microbiology and biotechnology , clostridium , biology , biochemistry , genetics , bacteria , neuraminidase , enzyme , computer science , programming language
Escherichia coli was transformed with pUC vectors containing Sau3A restriction fragments (RF) of Clostridium perfringens DNA. Two clones expressed sialidase activity when assayed with the fluorogenic substrate 4‐methylumbelliferyl‐α‐D‐ N ‐acetylneuraminic acid. A synthetic oligonucleotide representing the N‐terminus of the expressed enzyme hybridized with the clostridial insert and with a corresponding 2.1 kb Sau3A RF of the C. perfringens genome. The insert reduced to 1.4 kb, which still encoded active sialidase, has been sequenced. The structural gene encodes 382 amino acids representing an M r of 42 770. A hydrophobic leader sequence is absent. Upstream from the initiation codon ATG, a GA‐rich region is found and considered as the Shine‐Dalgarno sequence. Homology with the N‐terminus of the Vibrio cholerae sialidase gene and with viral sialidase sequences was not found.