Stable expression of the hepatitis B virus surface antigen containing pre-S2 protein in mouse cells using a bovine papillomavirus vector.
The large BglII fragment (2.8 kilobases) of hepatitis B virus DNA including the transcription unit for the hepatitis B surface antigen (HBsAg) was inserted into a bovine papillomavirus vector containing the neomycin resistance gene. The recombinant DNA was transfected into mouse C127 cells. A stable transformed cell line (MS128) secreting a large amount of 22 nm HBsAg particles containing pre-S2 protein was established. The secreted HBsAg particles had the receptor for polymerized human serum albumin. Immunoprecipitation and Western blot analyses showed that HBsAg particles consisted of two major proteins of 22K and 26K encoded by the S gene and a minor protein of 35K encoded by the pre-S2 and S genes. Southern blot analysis revealed that the transfected plasmid was integrated into the host chromosomal DNA and that most of the plasmid sequences were present. These results suggest that the stable expression of the HBsAg in MS128 cells is related to the integrated state of the recombinant DNA.
Yoneyama
T
T
Department of Enteroviruses, National Institute of Health, Tokyo, Japan.
Akatsuka
T
T
Miyamura
T
T
D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D002499:Centrifugation, Density Gradient; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D005814:Genes, Viral; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology; D007118:Immunoassay; D051379:Mice; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics; D010957:Plasmids; D011498:Protein Precursors / Q000235:genetics; D017293:Protein S; D014162:Transfection
D004274:DNA, Recombinant; D004279:DNA, Viral; D006023:Glycoproteins; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D017293:Protein S; C050930:protein S precursor