Document Detail

Expression of a cloned beta-glucanase gene from Bacillus amyloliquefaciens in an Escherichia coli relA strain after plasmid amplification.
MedLine Citation:
PMID:  3071739     Owner:  NLM     Status:  MEDLINE    
Amino acid starvation of cells of the Escherichia coli relA strain, CP79, which cannot accumulate guanosine tetraphosphate (ppGpp) in response to amino acid limitation, increased the pEG1 plasmid content about 5- to 7-fold in comparison with exponentially growing cells (pEG1:pBR322 with an insertion of Bacillus amyloliquefaciens DNA coding for beta-glucanase). In contrast, no pEG1 amplification occurred in E. coli CP78, the stringently controlled counterpart, after amino acid starvation. In order to verify these results, the plasmid DNA content was monitored by measuring the expression of pEG1-encoded beta-glucanase from B. amyloliquefaciens both before and after plasmid amplification. When amino acid starved CP79 cells were given an additional dose of amino acids, a more than 10-fold increase in pEG1-encoded beta-glucanase activity (per cell mass) was measured. This increase in enzyme activity correlates with pEG1 amplification during amino acid limitation. Under comparable conditions the activity of beta-glucanase was not increased in strain CP78, which did not amplify the plasmid. We suggest that the replication of pEG1 in amino acid starved E. coli cells is somehow under negative control by ppGpp. Moreover, we found the Bacillus beta-glucanase in E. coli relA cells to be excreted into the growth medium after starvation and overexpression.
M Hecker; S Riethdorf; C Bauer; A Schroeter; R Borriss
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Molecular & general genetics : MGG     Volume:  215     ISSN:  0026-8925     ISO Abbreviation:  Mol. Gen. Genet.     Publication Date:  1988 Dec 
Date Detail:
Created Date:  1989-05-19     Completed Date:  1989-05-19     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0125036     Medline TA:  Mol Gen Genet     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  181-3     Citation Subset:  IM    
Department of General Microbiology, Ernst-Moritz-Arndt-University, DDR, Greifswald.
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MeSH Terms
Amino Acids / pharmacology
Bacillus / enzymology,  genetics*
Cloning, Molecular
Escherichia coli / drug effects,  enzymology,  genetics
Gene Amplification
Gene Expression Regulation
Genes, Bacterial*
Glycoside Hydrolases / genetics*
Guanosine Tetraphosphate / genetics
Reg. No./Substance:
0/Amino Acids; 33503-72-9/Guanosine Tetraphosphate; EC 3.2.1.-/Glycoside Hydrolases; EC

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