Document Detail


Recombinant human and rat ciliary neurotrophic factors.
MedLine Citation:
PMID:  1861138     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The human ciliary neurotrophic factor (CNTF) gene was identified and cloned, based on homology with the recently cloned rat cDNA. The gene encodes a protein of 200 amino acids, which shares about 80% sequence identity with rat and rabbit CNTF and, like these homologues, lacks an apparent secretion signal sequence. The human CNTF gene, like the rat gene, appears to contain a single intron separating two protein coding exons. An intronless human CNTF gene was constructed by the use of polymerase chain reactions and introduced into vectors designed for expression of foreign proteins in E. coli. The rat CNTF gene was also introduced into similar vectors. Both the human and rat proteins were expressed at exceptionally high levels, at 20-40% and 60-70% of total protein, respectively. Extraction of the recombinant proteins from inclusion bodies by guanidinium chloride, followed by two column chromatography steps, produced high yields of pure CNTF that supported survival and neurite outgrowth from embryonic chick ciliary neurons in culture. The biological activity of both recombinant proteins was comparable to that of native rat CNTF.
Authors:
P Masiakowski; H X Liu; C Radziejewski; F Lottspeich; W Oberthuer; V Wong; R M Lindsay; M E Furth; N Panayotatos
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  57     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  1991 Sep 
Date Detail:
Created Date:  1991-09-04     Completed Date:  1991-09-04     Revised Date:  2006-05-01    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1003-12     Citation Subset:  IM    
Affiliation:
Regeneron Pharmaceuticals Inc., Tarrytown, New York.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Chromatography, Gel
Ciliary Neurotrophic Factor
Cloning, Molecular
DNA, Recombinant / genetics
Escherichia coli / genetics,  metabolism
Gene Expression
Genes / genetics
Genetic Vectors
Humans
Molecular Sequence Data
Nerve Tissue Proteins / genetics*,  metabolism
Plasmids
Rats
Recombinant Proteins / genetics
Chemical
Reg. No./Substance:
0/Ciliary Neurotrophic Factor; 0/DNA, Recombinant; 0/Nerve Tissue Proteins; 0/Recombinant Proteins

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


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