Cell-surface expression of influenza virus haemagglutinin in insect cells using a baculovirus vector

Area of Science:

• Molecular Biology
• Virology
• Biotechnology

Background:

• Baculovirus expression systems are widely used for protein production.
• Influenza virus haemagglutinin (HA) is a key surface glycoprotein involved in viral entry.

Purpose of the Study:

• To construct a baculovirus transfer vector for expressing influenza virus HA.
• To characterize the expression and localization of HA in insect cells.

Main Methods:

• Construction of a hybrid HA/polyhedrin gene.
• Generation of recombinant Autographa californica nuclear polyhedrosis virus (AcNPV).
• Analysis of HA expression, binding to monoclonal antibodies, haemadsorption assays, immunoprecipitation, and mRNA analysis.

Main Results:

• Polyhedrin-negative AcNPV recombinants expressing HA activity were obtained.
• Insect cell-derived HA was plasma membrane-bound and recognized by a specific monoclonal antibody.
• A 65 kDa protein was identified, and mRNA analysis revealed a major transcript of 2.95 kilobases.

Conclusions:

• Autographa californica nuclear polyhedrosis virus (AcNPV) can be utilized to express plasma membrane-bound virus glycoproteins.
• This approach facilitates the study of viral glycoproteins in a heterologous system.