Document Detail


Evidence for prokaryotic transcription and translation control regions in the human factor IX gene.
MedLine Citation:
PMID:  2965665     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A human factor IX cDNA clone isolated from a liver cDNA library constructed in phage lambda gt11 vector was shown to express factor IX protein in Escherichia coli. A factor IX immunospecific protein of 46.8 kDa was expressed, but was not a beta-galactosidase-factor IX fusion protein. Expression was seen when the factor IX cDNA was cloned into two different vector systems, lambda gt11 and pUC9, in both orientations with respect to the vector lacZ promoter. The expression of factor IX was not under control of the lacZ promoter of either vector system. In addition, when the factor IX cDNA fragment was subcloned in both orientations into a promoterless cloning vector (p CPP3), the factor IX cDNA fragment demonstrated promoter activity when inserted in only one orientation resulting in expression of chloramphenicol acetyl transferase in E. coli and Bacillus subtilis. A DNA computer search of the N-terminal sequences of the factor IX gene revealed prokaryotic-like promoter and ribosome-binding site (RBS) sequences with strong homology to the E. coli consensus sequences. The predicted sites homologous with prokaryotic promoter and RBS consensus sequences are followed by an in-frame methionine which could correspond to the translation start codon of the expressed factor IX. This report provides the first evidence that a eukaryotic gene encodes the information necessary for both transcription and translation to control gene expression in a prokaryotic host.
Authors:
P J Simpson; W G Chou; A A Whitbeck; F E Young; S Zain
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Gene     Volume:  61     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  1987  
Date Detail:
Created Date:  1988-05-24     Completed Date:  1988-05-24     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  373-83     Citation Subset:  IM    
Affiliation:
Miles Laboratories, Inc., Biotechnology Group, Elkhart, IN 46515.
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MeSH Terms
Descriptor/Qualifier:
Bacteriophage lambda / genetics
Base Sequence
Cloning, Molecular
DNA, Recombinant*
Escherichia coli / genetics
Factor IX / genetics*
Gene Expression Regulation
Humans
Immunochemistry
Liver
Molecular Sequence Data
Protein Biosynthesis*
Transcription, Genetic*
Grant Support
ID/Acronym/Agency:
HL-30616/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/DNA, Recombinant; 9001-28-9/Factor IX

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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