[The construction of reporter plasmid of ULBP1 and preliminary studying on the influence of NS3/4A on transcription of ULBP1].
OBJECTIVE
To construct a report vector of ULBP1 promoter gene and preliminary study on the influence of NS3/4A on transcription of ULBP1.


METHODS
ULBP1 core promoter sequence was amplified by PCR, and inserted into pGL3-enhance vector, constructing ULBP1 reporter plasmid pGL3-ULBP1; The HCV protease NS3/4A gene was subcloned into pcDNA3-Flag vector (pcDNA3-Flag-NS3/4A), and the expression of this plasmid was demonstrated by Western blot. The influence of inhibition by NS3/4A on the level of ULBP1 transcription was tested by assaying the Luciferase activity in cells transfected with pGL3-ULBP1 with Luminometer.


RESULTS
The reporter plasmid of ULBP1 promoter gene and the eukaryotic expression plasmid Flag-NS3/4A have been constructed and expressed successfully; and the protease NS3/4A inhibit the level of ULPB transcription.


CONCLUSIONS
The protease NS3/4A of HCV down-regulates ULBP1 expression by inhibiting the transcription of ULBP1.
Ma
Hong-fang
HF
Beijing Institute of Biotechnology, Beijing 100850, China.
He
Xiang
X
Wang
Ke
K
Wang
Fang
F
Jiao
Xin-an
XA
Zhang
Yan-hong
YH
Zhang
Jin-gang
JG
Zhong
Hui
H
D015153:Blotting, Western; D045744:Cell Line, Tumor; D058851:GPI-Linked Proteins; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D014158:Transcription, Genetic; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D058851:GPI-Linked Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008156:Luciferases; D008565:Membrane Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C423231:ULBP1 protein, human