[Secretory expression of sCD14 in Pichia pastoris and its LPS binding activity].
OBJECTIVE
To achieve secretory expression of mature sCD14 gene in Pichia pastoris and to analyze its LPS binding activity.


METHODS
sCD14 gene was cloned into yeast expression vector pPIC9K. Recombinant plasmid was lineared by Sac I and transformed into Pichia pastoris GS115. Positive integrated clones were screened and expressed by methanol inducing.


RESULTS
The mature sCD14 was expressed in BMMY with pH 6.5 after optimization of inducing expression condition. Western blot and ELISA data revealed recombinant products combined with specific antibody. The results from FACS showed recombinant product had biological activity of binding with LPS.


CONCLUSIONS
The recombinant mature sCD14 molecules with favorable LPS-binding activity could be used for further study on its structure and functions.
Hou
Ben-xiang
BX
Faculty of Stomatology, Capital University of Medical Sciences, Beijing 100050, China. bjbenxiang@21cn.net
Wang
Hui
H
Zhang
Chen
C
Yin
Jun
J
D012634:Bodily Secretions; D015870:Gene Expression; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics
D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides