HBV-related gene c18orf54 cloning and preliminary function

doi:10.3969/j.issn.1006-7795.2011.03.002

Journal of Capital Medical University ›› 2011, Vol. 32 ›› Issue (3): 318-323.doi: 10.3969/j.issn.1006-7795.2011.03.002

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HBV-related gene c18orf54 cloning and preliminary function

CHANG Lu-si¹, QIAO Yong¹, WEI Hong-lian², XIAO Fan¹, HAO Xiao-hua¹, ZHANG Ren-wen³, CHENG Jun¹, WEI Hong-shan^1*

1. Beijing Ditan Hospital Institute of Infectious Diseases, Capital Medical University, Beijing 100015, China;2. Department of , Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;3. Graduate School of Peking University, Peking University Health Science Center, Beijing 100191, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-06-21 Published:2011-06-21
Contact: WEI Hong-shan

Abstract

Abstract: Objective To express and purify the recombinant protein, and to prepare the c18orf54 specific rabbit polyclonal antibody.
Methods c18orf54 cDNA was ligated into the prokaryotic expressive vector pET-32a(+), and the resulting plasmid was transformed into E.coil BL21(DE3). The protein expression was induced with IPTG and the protein was analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting. The expressed product was purified using Ni⁺ affinity column chromatography. The recombinant protein was also confirmed with mass spectrometry. Then the purified C18ORF54 fusion protein was used to immunize New Zealand rabbits to gain polyclonal antibody. The specificity and potency of polyclonal antibody were evaluated by Western blotting and ELISA.
Results The C18ORF54 fusion protein was highly expressed. The protein was mainly in the inclusion body. ELISA indicated the titer of polyclonal antibody>1∶320 000. The high specificity was confirmed with Western blotting. Immunohistochemical staining of liver tissue showed that this glycosyltransferase was mainly scattered in cytoplasm of hepatocytes.
Conclusion The recombinant C18ORF54 fusion protein and the c18orf54 specific polyclonal antibody will be valuable tools for the investigation on the biological function of C18ORF54.

Key words: hepatitis B virus, c18orf54, cell cycle, pathogenesis

CLC Number:

R 512.6

CHANG Lu-si, QIAO Yong, WEI Hong-lian, XIAO Fan, HAO Xiao-hua, ZHANG Ren-wen, CHENG Jun, WEI Hong-shan. HBV-related gene c18orf54 cloning and preliminary function[J]. Journal of Capital Medical University, 2011, 32(3): 318-323.

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https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2011/V32/I3/318

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HBV-related gene c18orf54 cloning and preliminary function

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