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KMID : 0578319910010040497
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Molecules and Cells
1991 Volume.1 No. 4 p.497 ~ p.502
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Molecular Cloning of an Immunoglobulin Gene Encoding the Haevy Chain of a Monoclonal Antibody specific for the pre-S2 Antigen of Hepatitis B Virus
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Ryu, Chun Jeih
Jin, Byung Rae/Hong, Hyo Jeong/Han, Moon Hi
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Abstract
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We have cloned an immunoglobulin gene which encodes the heavy chain of a murine monoclonal antibody (mAb) (y2b, x) specific for the pre-S2 surface antigen of HBV from H8 hybridoma cell line. The genomic library, harbored in X EMBL3 and constructed from the H8 cells, was screened in parallel with the variable region of heavy chain-specific cDNA coding for the H8 mAb and the Ig heavy chain gene intron enhancer-specific oligonucleotide used as probes. One (M-1) out of 200,000 plaques gave positive hybridization signal with both probes. Southern analysis of the positive clone revealed that a 6.6-kb EcoRI fragment carries a functionally rearranged heavy chain variable region (VH) gene and the Ig heavy chain gene intron enhancer. The nucleotide sequence analysis showed that the gene contains a functionally rearranged VH gene and the transcriptional elements required for B cell-specific expression. The VH gene fused with human Cyl gene was expressed in murine myeloma cells, and directed the synthesis of the heavy chain mRNA, suggesting that the immunoglobulin gene is functional.
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