[Expression and characterization of human pulmonary surfactant-associated protein A1 in Saccharomyces cerevisiae].
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.
Lan
H K
HK
Pediatrics of Zhujiang Hospital, First Miltary Medical University, Guangzhou 510282, China. lanhk@263.net or hklan0@yahoo.com
Feng
Z C
ZC
Huang
J S
JS
Zheng
W Y
WY
Chen
L S
LS
Chunyu
L J
LJ
Ren
D M
DM
D018274:Electroporation; D006801:Humans; D010587:Phagocytosis; D011510:Proteolipids / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D037662:Pulmonary Surfactant-Associated Protein A / Q000031:analogs & derivatives*; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics
D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D011994:Recombinant Proteins; C445842:SFTPA1 protein, human