Chemical synthesis of a human interferon-alpha 2 gene and its expression in Escherichia coli.

Nucleic acids research ·Vol. 11 ·No. 18 ·1983-09-24 ·Pages 6419-35

Edge MD, Greene AR, Heathcliffe GR, Moore VE, Faulkner NJ, Camble R, Petter NN, Trueman P, Schuch W, Hennam J

Abstract

A 511-base pair DNA fragment encoding human interferon-alpha 2 has been chemically synthesised and expressed from a lac UV5 and a synthetic trp promoter in Escherichia coli. The synthesis involved preparation of 68 oligodeoxyribonucleotides and their enzymic ligation. The product expressed from the trp promoter system had high antiviral activity and displayed biological effects similar to those of Namalwa interferon on natural killer cell activity and in a Daudi cell growth inhibition assay. E.coli minicells containing plasmid DNA with the synthetic IFN-alpha 2 gene under trp promoter control produce a protein with the same electrophoretic mobility as a sample of authentic IFN-alpha 2. The protein from E.coli cross-reacts with the monoclonal antibody NK-2 and was readily purified, close to homogeneity, by immunoadsorption chromatography on NK-2 sepharose.

MeSH Terms

Amino Acid Sequence Base Sequence Biological Assay Cell Line Cloning, Molecular DNA Replication/drug effects DNA Restriction Enzymes Escherichia coli/genetics Humans Interferon Type I/chemical synthesis,genetics,pharmacology Kinetics Nucleic Acid Hybridization Oligonucleotides/chemical synthesis Plasmids

Chemicals

Interferon Type I Oligonucleotides DNA Restriction Enzymes

Authors & Affiliations

10 authors, click to expand affiliations / ORCID

Edge M D

Greene A R

Heathcliffe G R

Moore V E

Faulkner N J

Camble R

Petter N N

Trueman P

Schuch W

Hennam J

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/11.18.6419

Published

1983-09-24

Pages

6419-35

Language

English

Region

England

NLM ID

0411011

PMCID

PMC326383

Subset

Databases

GENBANK

X01628

External Links

PubMed source DOI: 10.1093/nar/11.18.6419 PMC Full Text View journal details

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Chemical synthesis of a human interferon-alpha 2 gene and its expression in Escherichia coli.

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