Screening of hepatocyte proteins binding with the middle surface protein of the hepatitis B virus by the yeast two‑hybrid system

Li,Zhiqun; Linghu,Enqiang; Cheng,Jun

doi:10.3892/mmr.2014.2069

June-2014 Volume 9 Issue 6

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June-2014 Volume 9 Issue 6

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Article

Screening of hepatocyte proteins binding with the middle surface protein of the hepatitis B virus by the yeast two‑hybrid system

Authors:
- Zhiqun Li
- Enqiang Linghu
- Jun Cheng
View Affiliations / Copyright

Affiliations: Department of Gastroenterology and Hepatology, Chinese PLA General Hospital, Beijing 100853, P.R. China, Institute of Infectious Diseases, Ditan Hospital, Capital Medical University, Beijing 100015, P.R. China
Pages: 2342-2346
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Published online on: March 24, 2014

https://doi.org/10.3892/mmr.2014.2069
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Abstract

The effect of the middle hepatitis B virus surface protein (MHBs) remains to be elucidated. To investigate the biological function of the MHBs protein, the present study performed yeast two‑hybrid screening to search for proteins that interact with the MHBs protein in hepatocytes. The bait plasmid expressing the MHBs protein was constructed by cloning the gene of the MHBs protein into pGBKT7, then the recombinant plasmid DNA was transformed into AH109 yeast (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing the liver cDNA library plasmid in 2X yeast peptone dextrose adenine (YPDA) medium. The mated diploid yeast was plated on quadruple dropout medium (SD/‑Trp‑Leu‑His‑Ade) containing X‑α‑gal for selection and screening. Following extracting and sequencing of the plasmids from positive (blue) colonies, the sequence analysis was conducted and analyzed by bioinformatics methods. Two colonies were selected and sequenced. Among them, one was the human DNA sequence from the clone RP11‑490D19 on chromosome 9 and the other was homo sapiens 12 BAC RP11‑180M15 (Roswell Park Cancer Institute Human BAC Library). The yeast two‑hybrid system is an effective method for identifying hepatocyte proteins that interact with MHBs. The MHBs protein binds with different proteins suggesting that it has multiple functions in vivo.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Screening of hepatocyte proteins binding with the middle surface protein of the hepatitis B virus by the yeast two‑hybrid system

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