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| Expression and Identification of Recombinant Human Apolipoprotein C-I in Pichia pastoris |
| SU Man-man1, CHEN Yue1, WANG Ding-ding2, XU Tian-min3, YAN Wei-qun 1 |
1. College of Pharmacy, Jilin University, Changchun 130021,China;
2. College of Life Science and Biopharmacology, Guangdong Pharmaceutical University, Guangzhou 510006,China;
3. The Second Hospital, Jilin University, Changchun 130041, China |
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Abstract Apolipoprotein C-I (ApoC-I) is a small, basic apolipoprotein which is mainly secreted by the liver as a component of triglyceride-rich lipoproteins and high density lipoproteins whose importance in plasma lipoprotein metabolism is increasingly evident. Therefore, a Pichia pastoris expression system was first used to obtain a high level expression of secreted, recombinant human ApoC-I (rhApoC-I). The full-length ApoC-I encoding sequence, gained by RT-PCR, was inserted into the pPICZαC vector and transform Pichia pastoris strain X33 after sequencing. High expression transformants with zeocin resistance were detected by SDS-PAGE and Western blot analysis. The antiproliferation effects of rhApoC-I on rat coronary artery smooth muscle cells was analysed by the methylthiazoletetrazolium (MTT) assay. It was showed that there were rhApoC-I(6.6kDa) in the culture supernatant induced by methanol. The expression level of rhApoC-I was 80mg/L. RhApoC-I can inhibit the proliferation which induced by PDGF remarkably. Ground on this, the composition and functions of rhApoC-I can be studied.
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Received: 19 April 2012
Published: 25 August 2012
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