Influence of Nucleotide Sequences Related to the 5'-untranslated Leader Region of Polyhedrin mRNA on the Expression of HBsAg in Insect Cells
Nai-Yueh Tien
Depertment of Zoology, National Taiwan University, Taipei, Taiwan 107, R.O.C.
Chu-Fang Lo
Depertment of Zoology, National Taiwan University, Taipei, Taiwan 107, R.O.C.
Pei-Lain Yang
Depertment of Biology, Fu-Jen Catholic University, Taipei, Taiwan 242, R.O.C.
Ting-Ting Wu
Depertment of Zoology, National Taiwan University, Taipei, Taiwan 107, R.O.C.
Ming-Der Yang
Depertment of Zoology, National Taiwan University, Taipei, Taiwan 107, R.O.C.
Chung-Hsiung Wang
Depertment of Plant Pathology and Entomology, National Taiwan University, Taipei, Taiwan 107, R.O.C.
Chao-Hung Lee
Depertment of Pathology, Indiana University School of Medicine, Indianapolis, IN 46223, USA
Malcolm J. Fraser
Depertment of Biological Science, University of Notre Dame, Notre Dame, IN 46556, USA
Guang-Hsiung Kou
Depertment of Zoology, National Taiwan University, Taipei, Taiwan 107, R.O.C.
A series of new baculovirus transfer vectors, under the control of the polyhedrin promoter for foreign gene expression, was constructed and utilized to successfully express the HBsAg gene in insect cells. The antigenicity of expressed HBsAg was confirmed by Western blot and immunoprecipitation.
Electron microscope was used to examine the morphology of 22-nm lipoprotein particles. Comparison of the levels of nonfused HBsAg proteins expressed by three differently constructed recombinant vectors was made using SDS-PAGE, quantitative analysis of enzyme-immunoassay, and RNA dot blot analysis of total cytoplasmic RNA. The results suggested, the nucleotide sequences related to the 5'-untranslated leader region of polyhedrin mRNA were important to HBsAg expression, and the HBsAg synthesis in insect cells driven by the polyhedrin promoter do not appear to follow the Kozak rule.
