Screening and cloning of the genes of protein interacting with the N-terminal protein of hepatitis B virus DNA polymerase by yeast-two hybrid technique / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 84-86, 2005.
Article
in Zh
| WPRIM
| ID: wpr-333041
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To screen and clone the genes of protein interacting with the N-terminal protein (TP) of hepatitis B virus DNA polymerase.</p><p><b>METHODS</b>TP was amplified by polymerase chain reaction (PCR) and TP bait plasmid was constructed by using yeast two-hybrid system 3, then transformed into yeast AH 109. The transformed yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast was plated on synthetic dropout medium (SD/-Trp-Leu-His-Ade) and that containing X-alpha-GAL for selecting two times and screening. Plasmids were extracted from blue colonies, and sequence analysis was performed by bioinformatics.</p><p><b>RESULTS</b>Forty-seven clones were obtained, these clones included human P36956 sterol regulatory element binding protein-1, RNA polymerase II subunit hsRPB7 mRNA, asialoglycoprotein receptor 2, transcript variant 3, ceruloplasmin (ferroxidase), transmembrane 4 superfamily member 2 and 19 of the hypothetical proteins and so on.</p><p><b>CONCLUSION</b>Genes encoding TP interacting proteins in hepatocytes were successfully cloned and the results suggest that TP has a wide variety of biological functions.</p>
Full text:
1
Index:
WPRIM
Main subject:
Plasmids
/
Protein Binding
/
Receptors, Virus
/
Transformation, Genetic
/
Viral Proteins
/
Gene Library
/
Chemistry
/
Hepatitis B virus
/
Cloning, Molecular
/
Two-Hybrid System Techniques
Type of study:
Diagnostic_studies
/
Screening_studies
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2005
Type:
Article