Cloning, Sequence, and Expression of the L-(+) Lactate Dehydrogenase of Streptococcus bovis |
| |
Authors: | HA Wyckoff J Chow TR Whitehead MA Cotta |
| |
Institution: | (1) Fermentation Biochemistry Research Unit, National Center for Agricultural Utilization Research, U.S. Department of Agriculture, Agricultural Research Service, 1815 N. University Street, Peoria, IL 61604, USA , US |
| |
Abstract: | The ldh gene encoding the
fructose-1,6-diphosphate-dependent L-(+) lactate dehydrogenase from the
ruminal bacterium Streptococcus bovis was cloned and sequenced. A
genomic library of S. bovis JB1 DNA was constructed in lambda ZAP II
and screened by use of a heterologous probe derived from the cloned
Streptococcus mutans ldh gene. Several clones were isolated that
contained a common 2.9-kb fragment as determined by restriction analysis.
Nucleotide sequence analysis revealed a 987-bp open reading frame with
extensive homology to Streptococcus thermophilus and S. mutans
ldh nucleic acid and amino acid sequences. Expression of the cloned
S. bovis ldh gene in Escherichia coli was confirmed by the
ability to complement the ldh mutation of E. coli FMJ39, by
using an in-gel activity screen and by enzymatic assay. Increased LDH
activity was observed in S. bovis JB1 containing the cloned
ldh genes on a multicopy plasmid.
Received: 15 October 1996 / Accepted: 3 December 1996 |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|