| Summary: | 碩士 === 中山醫學院 === 生物化學研究所 === 84 === Interleukin 13 (IL-13)is a recently found cytokine secreted
by activated T cells that is a potent in vitro modulator of
human monocyteand B cell functions. The studies of IL-13 and
IL-4 indicate that they share many common biological activites.
Both IL-13 and IL-4 inhibit the production of
proinflammatorycytokines, chemokines and hemotopoietic growth
factorsby human monocyte, and can also enhance the Ig isotype
class switching and the secretion of IgE and IgG4. These
indicate that IL-13 play an important role in anti-inflammatory
responses, allergic response and many clinical diseases. In
order to further investigate the functions and roles of hIL-13,
we hope to obtain a large quantify and reliable source of
hIL-13. Firstly, we use a E. coli/pET-30 expression system to
produce recombinant hIL-13 protein. hIL-13 DNA wsa ligated with
pET-30 plasmid and transformed them into E. coli BL21 (DE3).
hIL-13 fusion protein was purified using His-Binding Resin.
Since hIL-13 is an eukaryotic protein, we also use a
Baculovirus/insect cell system to express hIL-13. Two transfer
plasmids (pVL1393, pAcG2T) was ligatedwith hIL-13 cDNA,
respectively, and co-transfected the ligands with Baculovirus
DNA into SF-9 inscet cell line. In this eukaryotic system, we
wish to obtain recombinant hIL-13 that will resemble its native
confer part. The biological activity of the recombinant hIL-13
was determined by using TF-1 cell line and human monocytes.
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