PMID- 16261208
OWN - NLM
STAT- MEDLINE
DCOM- 20081222
LR  - 20051101
IS  - 1003-9279 (Print)
IS  - 1003-9279 (Linking)
VI  - 19
IP  - 3
DP  - 2005 Sep
TI  - [Screening and cloning of hepatitis C virus non-structural protein 4B interacting 
      protein gene in hepatocytes].
PG  - 248-51
AB  - OBJECTIVE: To investigate biological functions of non-structural protein 4B 
      (NS4B) of hepatitis C virus (HCV), yeast-two hybrid technique was performed to 
      seek proteins in hepatocytes interacting with HCV NS4B. METHODS: HCV NS4B bait 
      plasmid was constructed by ligating the NS4B gene with carrier plasmid pGBKT7 and 
      transformed into yeast cells AH109 (type alpha). The transformed yeast cells were 
      amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA 
      library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on 
      synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout 
      nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two 
      times. After extracting plasmid from blue colonies, plasmid DNA was transformed 
      into competent Escherichia coli and analysed by DNA sequencing and 
      bioinformatics. RESULTS: Five genes in eight positive colonies were obtained. 
      There were one NADH dehydrogenase subunit 3, one cytochrome c oxidase subunit 
      III, one retinol binding protein 4, one reticulon 3-A (RTN3) and one fibrinogen 
      gamma polypeptide (FGG). CONCLUSION: Genes of HCV NS4B interacting proteins in 
      hepatocytes were successfully cloned and the results paved the way for studying 
      the biological functions of NS4B and associated proteins.
FAU - Liu, Yan
AU  - Liu Y
AD  - Gene Therapy Research Center, Institute of Infectious Diseases, The No. 302 
      Hospital of the People's Liberation Army, Beijing 100039, China.
FAU - Cheng, Jun
AU  - Cheng J
FAU - Bai, Gui-qin
AU  - Bai GQ
FAU - Yan, Fu-ming
AU  - Yan FM
FAU - Wu, Shun-hua
AU  - Wu SH
FAU - Wang, Lin
AU  - Wang L
FAU - Zhang, Ling-xia
AU  - Zhang LX
LA  - chi
PT  - English Abstract
PT  - Journal Article
PL  - China
TA  - Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
JT  - Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang 
      bingduxue zazhi = Chinese journal of experimental and clinical virology
JID - 9602873
RN  - 0 (Carrier Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (RTN3 protein, human)
RN  - 0 (Retinol-Binding Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - EC 1.6.99.3 (NADH Dehydrogenase)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Blotting, Western
MH  - Carrier Proteins/genetics/metabolism
MH  - Cell Line, Tumor
MH  - Cloning, Molecular
MH  - Electron Transport Complex IV/genetics/metabolism
MH  - Gene Library
MH  - Hepatocytes/*metabolism
MH  - Humans
MH  - Immunoprecipitation
MH  - Membrane Proteins/genetics/metabolism
MH  - NADH Dehydrogenase/genetics/metabolism
MH  - Nerve Tissue Proteins/genetics/metabolism
MH  - Plasmids/genetics
MH  - Protein Binding
MH  - Retinol-Binding Proteins/genetics/metabolism
MH  - *Two-Hybrid System Techniques
MH  - Viral Nonstructural Proteins/genetics/*metabolism
EDAT- 2005/11/02 09:00
MHDA- 2008/12/23 09:00
CRDT- 2005/11/02 09:00
PHST- 2005/11/02 09:00 [pubmed]
PHST- 2008/12/23 09:00 [medline]
PHST- 2005/11/02 09:00 [entrez]
PST - ppublish
SO  - Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Sep;19(3):248-51.