[Expression and characterization of human pulmonary surfactant-associated protein A1 in Saccharomyces cerevisiae].
2001
The cDNA encoding pulmonary surfactant-associated protein A1 (SP-A1) derived from healthy adult's lung was cloned into the pVT102U/alpha, expression vector of Saccharomyces cerevisiae, which contains the yeast alpha-factor signal sequence, leading to the secretion of expressed protein, and then transformed into Saccharomyces cerevisiae S-78 (leu2, ura3, rep4) by electroporation. After 2-3 days culture in adequate pH, the expressed SP-A1 accumulated up to 400 mg/L in supernatant. The pure proteins were obtained by Sephadex G-25, G-75, Sepharose 4B. The expressed recombinant products, 62 kD and 32 kD, reacted to specific antibody using ELISA and Western blot. The SP-A1 protein expressed in Saccharomyces cerevisiae was efficient in enhancing the phagocytosis of E. coli J5 by alveolar macrophages.
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