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Updated: Jun 6, 2026
A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
[Expression, purification and antibody preparation of recombinant human SM22alpha]
Jin-kun Wen1, Lei Nie, Jing-yi Li
1Key Laboratory for Medical Biotechnology of Hebei Province, Institute of Basic Medical Sciences, Hebei Medical University, Shijiazhuang 050017, China. wjk@hebmu.edu.cn
Aim:
The recombinant human smooth muscle 22 alpha (SM22alpha) was expressed by using Pichia pastoris.
Methods:
Using pGEM3z-SM22alpha as the template, SM22alpha coding region was amplified by PCR, and was inserted the expression vector pPIC9. Then the recombinant plasmid pPIC9-SM22alpha was transfected into Pichia pastoris. The products induced by methanol were precipitated by ammonium sulfate, then CM-cellulose chromatography was performed for SM22alpha. Polyclonal antibody against SM22alpha was produced by immunizing a rabbit with purified recombinant SM22alpha.
Results:
The positive clone with SM22alpha got high output at 84 hours after induction by methanol. The SM22alpha prepared by ammonium sulfate fractionation and chromatographic separation showed a single band whose apparent molecular weight was 22 kD on SDS-PAGE. Polyclonal antibody against SM22alpha could detect the SM22alpha expression in human or rat vascular walls.
Conclusion:
High-level expression of SM22alpha is successfully achieved in Pichia pastoris. Antibody against SM22alpha can be used to explore the function of SM22alpha.
