Expression of human interleukin-11 in cell culture and larvae of silkworm.
- Author:
Xi-Jie GUO
1
;
Yong-Feng JIN
;
Ming-Guan YANG
;
Yao-Zhou ZHANG
Author Information
1. Institute of Biochemistry, Zhejiang University, Hangzhou 310029, China. guoxijie@fm365.com
- Publication Type:Journal Article
- MeSH:
Animals;
Baculoviridae;
genetics;
Bombyx;
cytology;
growth & development;
metabolism;
Cell Culture Techniques;
Gene Expression;
Genetic Vectors;
Humans;
Interleukin-11;
biosynthesis;
genetics;
Larva;
genetics;
metabolism;
Pupa;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2002;18(1):30-34
- CountryChina
- Language:Chinese
-
Abstract:
A recombinant transfer vector, pBacIL-11, containing hIL-11 cDNA of 546 nucleotides lacking leader sequence was constructed and co-transfected into BmN cells with linearized BmBacPAK(modified BmNPV) DNA for construction of a recombinant baculovirus carrying the hIL-11 gene. Southern hybridization analysis suggested that the recombinant baculovirus DNA contained hIL-11 cDNA fragment. RNA dot blotting demonstrated that the hIL-11 gene was transcribed. The recombinant baculovirus has a strong infectivity to BmN cell line and to silkworm larvae and pupae. Specific hIL-11 bands were detected from all the samples of cell extract, culture supernatant, haemolymph of larvae and pupae by SDS-PAGE analysis. Biological activity of the expressed product was determined with IL-11 dependent B9-11 cell line and by MTT colorimetric assay, which indicated that biologically active rhIL-11 protein was overexpressed in BmN cell line and in silkworm larvae and pupae.
