[Liver tissue-specific stable expression of human CD81 molecule]

Area of Science:

• * Molecular Biology
• * Virology
• * Cell Biology

Background:

• * Hepatitis C virus (HCV) entry into host cells is mediated by host factors, including CD81.
• * Understanding the role of human CD81 in HCV infection is crucial for developing antiviral strategies.

Purpose of the Study:

• * To establish a stable expression system for the human CD81 gene in a mouse hepatoma cell line (Hepa 1-6).
• * To utilize a liver-specific promoter for targeted gene expression in Hepa 1-6 cells.

Main Methods:

• * Isolation of human CD81 gene from HepG2 cells and RT-PCR amplification.
• * Ligation of liver-specific albumin promoter/enhancer and SV40 polyA sequence to the CD81 gene.
• * Construction of a recombinant eukaryotic expression vector (pcDNA3-Alb p-CD81).
• * Transfection of the recombinant vector into Hepa 1-6 cells and confirmation of gene expression via RT-PCR and FACS.

Main Results:

• * Successful cloning and sequencing of the human CD81 gene.
• * Stable transcription of human CD81 mRNA confirmed in transfected Hepa 1-6 cells.
• * Stable expression of human CD81 protein on the surface of Hepa 1-6 cells demonstrated.

Conclusions:

• * Established Hepa 1-6 cell clones stably expressing human CD81, a receptor for HCV.
• * Provides a foundation for studying HCV envelope protein interactions with human CD81.
• * Facilitates research on screening for HCV infection blocking drugs and developing HCV infection mouse models.