Secretion of the hepatitis B virus surface antigen from mouse cells using an extra-chromosomal eucaryotic vector

Stenlund, A., Lamy, D., Moreno-Lopez, J., Ahola, H., Pettersson, U., Tiollais, P. (1983) Secretion of the hepatitis B virus surface antigen from mouse cells using an extra-chromosomal eucaryotic vector. EMBO J, 2 (5). pp. 669-73. ISSN 0261-4189 (Print)0261-4189 (Linking)

URL: http://www.ncbi.nlm.nih.gov/pubmed/6641713

Abstract

Recombinant DNA molecules which contained a subgenomic fragment of the hepatitis B virus (HBV) genome, the pML2 vector and the bovine papillomavirus type 1 (BPV) genome were constructed. The HBV fragment includes the entire transcription unit for the hepatitis B surface antigen (HBsAg). After propagation in Escherichia coli, the recombinant plasmids were cleaved with endonucleases SalI and PvuI to eliminate most of the bacterial sequences before transfection of mouse C127 cells. Foci were observed 10--14 days after transfection. Cells from selected foci were cloned and the supernatants were assayed for the presence of HBsAg. Most of the clones tested were found to secrete HBsAg particles into the growth medium. These particles appear to be similar to the 22 nm particles present in the serum of HBV chronic carriers. SDS-polyacrylamide gel electrophoresis revealed that the particles contain two polypeptides, probably representing the glycosylated and unglycosylated forms of the HBsAg major polypeptide. An analysis of DNA from the transformed clones revealed that they contain multiple extra-chromosomal copies of the recombinant, which, however, had suffered rearrangement.

Item Type: Paper
Uncontrolled Keywords: Animals Base Sequence DNA, Viral/analysis Extrachromosomal Inheritance Hepatitis B Surface Antigens/ biosynthesis Hepatitis B virus/ genetics Mice RNA, Viral/analysis Replicon
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA replication
organism description > animal > mammal > rodent > mouse
organism description > virus
CSHL Authors:
Communities: CSHL labs > Stenlund lab
Depositing User: Matt Covey
Date: 1983
Date Deposited: 12 Mar 2013 18:39
Last Modified: 12 Mar 2013 18:39
PMCID: PMC555167
Related URLs:
URI: http://repository.cshl.edu/id/eprint/27781

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