Cloning and expression of canine interferon-gamma gene in mouse SP2/0 cell line / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 365-368, 2002.
Article
in Chinese
| WPRIM
| ID: wpr-256201
ABSTRACT
Canine Interferon-gamma (CaIFN-gamma) cDNA was cloned from spleen T cells of dog by reverse transcription-polymerase chain reaction (RT-PCR). CaIFN-gamma cDNA were digested with Hind III and Not I, and inserted into pRc/CMV2 expression vector. The pRc/CMV2/CaIFN-gamma vector was sequenced, and predicted to produce a signal peptide of 23 amino acids and a mature protein of 143 amino acids with a molecular weight of 19 kD. Two potential N-glycosylation sites are located at positions 16 and 83 of the mature protein. Comparison of the CaIFN-gamma protein sequence with that of CaIFN-gamma reported from DDBJ/GenBank revealed a homology of 99%. To establish a long time expression system, pRc/CMV2/CaIFN-gamma vector was transfected into mouse SP2/0 cell line. The SP2/0 cells culture supernatants was harvested and the antiviral activity was measured following cytopathic-effect inhibition assay using Madin-Darby Canine Kidney (MDCK)-vesicular stomatitis virus(VSV) system. Initial transformants with G418 phenotype produced recombinant CaIFN-gamma titers ranging from 2,500 to 5,000 u/mL of culture medium.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Recombinant Proteins
/
Molecular Sequence Data
/
Base Sequence
/
Cell Line
/
Chemistry
/
Amino Acid Sequence
/
Interferon-gamma
/
Cloning, Molecular
/
DNA, Complementary
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
Similar
MEDLINE
...
LILACS
LIS