Document Detail

Increased synthesis in E. coli of fibroblast and leukocyte interferons through alterations in ribosome binding sites.
MedLine Citation:
PMID:  6187521     Owner:  NLM     Status:  MEDLINE    
Eleven chimeric plasmids have been constructed which direct the synthesis of mature human fibroblast (IFN-beta 1) or leukocyte interferon (IFN-alpha A) proteins under the control of the E. coli trp promoter. The plasmids differ with respect to the nucleotide spacing between the Shine-Dalgarno sequence of the trp leader and the ATG translation start signal of the interferon genes. By utilizing a unique Xba I endonuclease site located within the spacer region of the expression plasmids, the spacings were altered from 2-10 nucleotides or 7-15 nucleotides for the fibroblast and leukocyte interferon expression plasmids, respectively. The optimal spacing for expression, as determined by interferon assay, is 9 nucleotides for both types of transcripts, despite differences in nucleotide sequence within the spacer region and downstream from the AUG initiator. Yields of IFN-alpha A varied about six-fold, while among the different IFN-beta 1 expression plasmids a range of more than 100-fold in interferon production was observed. The difference in the range of variation between the IFN-alpha A and IFN-beta 1 plasmids is attributed partly to changes in messenger RNA secondary structure within the ribosome binding sites which affect message half-life.
H M Shepard; E Yelverton; D V Goeddel
Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  DNA (Mary Ann Liebert, Inc.)     Volume:  1     ISSN:  0198-0238     ISO Abbreviation:  DNA     Publication Date:  1982  
Date Detail:
Created Date:  1983-05-27     Completed Date:  1983-05-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8302432     Medline TA:  DNA     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  125-31     Citation Subset:  IM    
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MeSH Terms
Binding Sites
Escherichia coli / genetics,  metabolism*
Interferon Type I / biosynthesis*,  genetics
Nucleic Acid Conformation
Protein Biosynthesis
RNA, Bacterial / metabolism
RNA, Messenger / metabolism
Ribosomes / metabolism*
Reg. No./Substance:
0/Interferon Type I; 0/RNA, Bacterial; 0/RNA, Messenger

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