Abstract：
:With primers designed for the conserved region of the gamma-butyrolactone autoregulator receptor proteins from Streptomyces species, PCR using the Streptomyces clavuligerus genome DNA as a template gave a clear band of 100 bp, the sequence of which revealed high similarity to the expected region of a receptor gene. By Southern blot and colony hybridization with the 100-bp insert as a probe, plasmid pSCA, harboring a 4.2 kb- SalI fragment, was obtained. Sequence analysis on the insert revealed a 702-bp ORF encoding a protein with a moderate similarity (identity, 33-43%; similarity, 51-62%) to known gamma-butyrolactone autoregulator receptor proteins from Streptomyces sp. The ORF was named scaR ( S. clavuligerus autoregulator receptor). The scaR/pET-3d plasmid was constructed for overexpression of the recombinant ScaR protein (rScaR) in Escherichia coli, and the rScaR protein was purified to homogeneity by DEAE-ion-exchange HPLC. The molecular mass of the purified rScaR protein was determined to be 27 kDa as determined by SDS-PAGE, and 54 kDa by gel filtration HPLC under nondenatured conditions at a low protein concentration, indicating that the majority of the native ScaR is present in the form of a dimer, although rScaR tended to aggregate into a higher molecular form of 230 kDa at a high protein concentration. A binding assay with tritium-labeled autoregulators indicated that IM-2 type compounds with a long C2 side chain were the most effective ligands for rScaR, demonstrating for the first time that the beta-lactam producer S. clavuligerus contains a gene for the gamma-butyrolactone autoregulator receptor.
journal_name
Arch Microbioljournal_title
Archives of microbiologyauthors
Kim HS,Lee YJ,Lee CK,Choi SU,Yeo SH,Hwang YI,Yu TS,Kinoshita H,Nihira Tdoi
10.1007/s00203-004-0697-xkeywords:
subject
Has Abstractpub_date
2004-09-01 00:00:00pages
44-50issue
1eissn
0302-8933issn
1432-072Xjournal_volume
182pub_type
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journal_title：Archives of microbiology
pub_type： 杂志文章
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pub_type： 杂志文章
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pub_type： 杂志文章
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journal_title：Archives of microbiology
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journal_title：Archives of microbiology
pub_type： 杂志文章
doi：10.1007/BF00427847
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pub_type： 杂志文章
doi：10.1007/BF00427869
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更新日期：2010-02-01 00:00:00
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pub_type： 杂志文章
doi：10.1007/s00203-013-0911-9
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pub_type： 杂志文章
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pub_type： 杂志文章
doi：10.1007/BF00406167
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doi：10.1007/BF00415614
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journal_title：Archives of microbiology
pub_type： 杂志文章
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更新日期：1997-04-01 00:00:00
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journal_title：Archives of microbiology
pub_type： 杂志文章
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