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[cDNA cloning and prokaryotic expression of human cholesteryl ester transfer protein and preparation of its antiserum].
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- Author(s): Liu HR;Liu HR; Zeng WW; Zhou B; Shen L; Wu G; Xue H; Chen BS
- Source:
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2008 May; Vol. 24 (5), pp. 471-4.
- Publication Type:
Journal Article
- Language:
Chinese
- Additional Information
- Source:
Publisher: Xi bao yu fen zi mian yi xue za zhi bian ji bu Country of Publication: China NLM ID: 101139110 Publication Model: Print Cited Medium: Print ISSN: 1007-8738 (Print) Linking ISSN: 10078738 NLM ISO Abbreviation: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi Subsets: MEDLINE
- Publication Information:
Original Publication: Xiʹan : Xi bao yu fen zi mian yi xue za zhi bian ji bu
- Subject Terms:
- Abstract:
Aim: To clone human cholesteryl ester transfer protein (CETP) cDNA, express and purify human CETP in E.coli and prepare CETP-specific rabbit antiserum.
Methods: by RT-PCR method, the encoding sequence of human cholesteryl ester transfer protein (CETP) was cloned into pET-30b(+) vector. Then BL21 (DE3) of E.coli transformed with recombinant vector pET-CETP was induced to express CETP in high level by IPTG. The expressed protein was purified from SDS-polyacrylamide gel, and the antiserum against CETP was raised in rabbit. The titer and specificity of rabbit antiserum were evaluated by ELISA, Western blot and immunofluorescence assay.
Results: The results of SDS-PAGE showed that CETP was expressed in the form of inclusion bodies in BL21(DE3) and the best expression time was about 4 hours. The titer of the rabbit antiserum prepared with CETP purified from SDS-PAGE was 1:5. 12 x 10(5) and the antiserum reacted specifically with CETP expressed in BL21 (DE3) and COS7 cells.
Conclusion: The preparation of the specific rabbit antiserum against CETP will be valuable for the study on the structure and function of human CETP.
- Accession Number:
0 (Antibodies, Monoclonal)
0 (CETP protein, human)
0 (Capsid Proteins)
0 (Cholesterol Ester Transfer Proteins)
0 (DNA, Complementary)
0 (Recombinant Proteins)
367-93-1 (Isopropyl Thiogalactoside)
- Publication Date:
Date Created: 20080510 Date Completed: 20100430 Latest Revision: 20201209
- Publication Date:
20221213
- Accession Number:
18466705
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