Isolation and expression of rat interferon beta gene and growth-inhibitory effect of its expression on rat glioma cells.
Rat interferon beta (IFN beta) was cloned from a part of rat genomic DNA by the polymerase chain reaction (PCR) with reference to the nucleotide sequence data of a mouse IFN beta. The sequence result showed that the clone had 555 bp of open reading frame (ORF) which encoded a 184-amino acid polypeptide. Comparison of our data with those of mouse and human IFN beta revealed 86.7 and 67.6% homology at the nucleotide level, and 76.2 and 48.9% homology at the amino acid level, respectively. This ORF was inserted into an expression vector, pCAGGS, and COS-1 cells were then transfected with the plasmid encapsulated in cationic multilamellar liposomes. From the results of reverse transcription-PCR and cytopathic activity assay, this ORF expressed IFN beta in COS-1 cells. When the cultured cells of rat glioma cell line T9 were transfected with the plasmid, their growth was markedly suppressed due to the expression of rat IFN beta.
Yokoyama
S
S
Institute of Applied Biochemistry, Gifu, Japan.
Ohishi
N
N
Shamoto
M
M
Watanabe
Y
Y
Yagi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002455:Cell Division; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005910:Glioma / Q000235:genetics / Q000473:pathology; D006131:Growth Inhibitors / Q000235:genetics*; D006801:Humans; D016899:Interferon-beta / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured
D006131:Growth Inhibitors; D016899:Interferon-beta; D004247:DNA