Document Detail

Transcriptional antitermination activity of the synthetic nut elements of coliphage lambda. I. Assembly of the nutR recognition site from boxA and nut core elements.
MedLine Citation:
PMID:  3005108     Owner:  NLM     Status:  MEDLINE    
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.
A L Brown; W Szybalski
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Gene     Volume:  39     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  1985  
Date Detail:
Created Date:  1986-04-03     Completed Date:  1986-04-03     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  121-7     Citation Subset:  IM    
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MeSH Terms
Bacterial Proteins / genetics
Bacteriophage lambda / genetics*
Base Sequence
DNA Restriction Enzymes / metabolism
Gene Expression Regulation
Genes, Regulator*
Genes, Viral*
Oligodeoxyribonucleotides / chemical synthesis
Transcription, Genetic*
Grant Support
5-P01-CA-23076/CA/NCI NIH HHS; 5-P30-CA-07175/CA/NCI NIH HHS; CA-09075/CA/NCI NIH HHS
Reg. No./Substance:
0/Bacterial Proteins; 0/Oligodeoxyribonucleotides; EC 3.1.21.-/DNA Restriction Enzymes

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