| Transcriptional antitermination activity of the synthetic nut elements of coliphage lambda. I. Assembly of the nutR recognition site from boxA and nut core elements. | |
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MedLine Citation:
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PMID: 3005108 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An active nutR antiterminator was reconstructed from two synthetic modules, one containing the 8-bp boxA (5'-CGCTCTTA) and the other the 17-bp nutR core (5'-AGCCCTGAAAAAGGGCA) sequence. The modules were synthesized with HindIII cohesive ends, which upon annealing and ligation created an 8-bp spacer (5'-CAAAGCTT) between the boxA and nutR core. The 8-bp length was the same as in the native nutR (5'-CACATTCC), but the sequence showed less than 38% homology. The antitermination mediated by the synthetic nutR, was 68-80% efficient when tested in the pp-nutR-N-tL1-galK expression plasmid, analogous to that used by Drahos and Szybalski [Gene, 16 (1981) 261-274]. The cloned boxA by itself has no activity, while the nutR core alone shows only marginal (5-10%) antiterminator function. Increasing the distance between boxA and the nutR core from 8 bp to 20-28 bp, i.e., by one to two turns of the DNA helix (about 10 bp per turn), has little effect on the antiterminator function, whereas use of spacers with length about halfway between 8 and 20 bp results in reduced antitermination. It appears that both the sequences and spacial arrangement of the boxA and nut elements are important for efficient antiterminator function. |
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Authors:
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A L Brown; W Szybalski |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Gene Volume: 39 ISSN: 0378-1119 ISO Abbreviation: Gene Publication Date: 1985 |
Date Detail:
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Created Date: 1986-04-03 Completed Date: 1986-04-03 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 7706761 Medline TA: Gene Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 121-7 Citation Subset: IM |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/M15218 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Bacterial Proteins
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genetics Bacteriophage lambda / genetics* Base Sequence DNA Restriction Enzymes / metabolism Gene Expression Regulation Genes, Regulator* Genes, Viral* Oligodeoxyribonucleotides / chemical synthesis Plasmids Transcription, Genetic* |
| Grant Support | |
ID/Acronym/Agency:
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5-P01-CA-23076/CA/NCI NIH HHS; 5-P30-CA-07175/CA/NCI NIH HHS; CA-09075/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/Oligodeoxyribonucleotides; EC 3.1.21.-/DNA Restriction Enzymes |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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