Expression of human interleukin-6 (IL-6) in an insect system

A cDNA fragment coding human interleukin-6 (IL-6) was inserted into the vector pSXIVVI +X3 with the control of Syn and XIV promoters. The Sf9 cells ( Spodoptera frugiperda) were co-transfected with the recombinant plasmid and TnNPV DNA ( Trichoplusia ni nuclear polyhedrosis virus DNA). Cells infecte...

Full description

Saved in:
Bibliographic Details
Published in:Bioresource technology 1999-05, Vol.68 (2), p.201-203
Main Authors: Zhang, Xue-Wu, Sun, Tao, Zhou, Xiao-Feng, Liu, Xin, Gu, De-Xiang, Huang, Xiao-Ni
Format: Article
Language:English
Subjects:
Antigens
Antineoplastic drugs. Cytokines
baculovirus expression vector system
baculovirus system
Bioassay
Biological and medical sciences
Biosynthesis
Biotechnology
cell lines
Colorimetry
complementary DNA
DNA
Extraction
Fundamental and applied biological sciences. Psychology
gene expression
genetic vectors
Health. Pharmaceutical industry
Industrial applications and implications. Economical aspects
interleukin-6
larvae
Molecular weight
Nucleopolyhedrovirus
Plusia
plusia agnata
Production of active biomolecules
Proteins
sf9 cells
Spodoptera frugiperda
transfer vector
Trichoplusia ni
Viruses
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A cDNA fragment coding human interleukin-6 (IL-6) was inserted into the vector pSXIVVI +X3 with the control of Syn and XIV promoters. The Sf9 cells ( Spodoptera frugiperda) were co-transfected with the recombinant plasmid and TnNPV DNA ( Trichoplusia ni nuclear polyhedrosis virus DNA). Cells infected with recombinant virus synthesized IL-6 protein at a level of approx. 42% of total cellular protein. IL-6 activity in infected cells was measured by using the IL-6-dependent mouse B hybridoma cell line 7TD1 and MTT colorimetric assay, the highest expression level, 2.82 × 10 6 u/ml, was obtained at 96 h after infection. Western blot analysis of infected larvae protein extracts showed that the virus-mediated IL-6, with a molecular weight of ∼21 kDa, shared antigenic sites with the authentic counterpart. However, no evidence of its bioactivity was found, IL-6 activity was determined only for IL-6 expressed by insect cells.
ISSN:0960-8524
1873-2976
DOI:10.1016/S0960-8524(98)00129-1