The use of consensus sequence for amplification and oriented cloning of human alpha interferon genes.
A clone from human cDNA library was amplified in polymerase chain reaction (PCR) by the synthetic oligonucleotides. The final construct after linker ligation and digestion with restriction endonucleases was suitable for oriented cloning into E. coli expression vector. The interferon (IFN) expression could be detected by SDS-PAGE electrophoresis. Western blotting and biological antiviral assay. This set of oligonucleotides can be used also for amplification of genomic DNA or cDNA libraries.
Kúdela
O
O
Institute of Virology, Slovak Academy of Sciences, Bratislava.
Zelník
V
V
Kaluz
S
S
Pastorek
J
J
D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D004247:DNA