[Expression of human interleukin-11 in cell culture and larvae of silkworm].
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.
Guo
Xi-Jie
XJ
Institute of Biochemistry, Zhejiang University, Hangzhou 310029, China. guoxijie@fm365.com
Jin
Yong-Feng
YF
Yang
Ming-Guan
MG
Zhang
Yao-Zhou
YZ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D012831:Bombyx / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D018929:Cell Culture Techniques; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D017370:Interleukin-11 / Q000096:biosynthesis* / Q000235:genetics; D007814:Larva / Q000235:genetics / Q000378:metabolism*; D011679:Pupa / Q000235:genetics / Q000378:metabolism
D017370:Interleukin-11