Synthesis of part of a mouse immunoglobulin light chain in a bacterial clone
| Autor: | D. Salomon, O. Amster, Israel Schechter, O. Zemel, Frida Kantor, Ada Zamir, Elisha Zeelon |
|---|---|
| Rok vydání: | 1980 |
| Předmět: |
Oligonucleotide
Genes MHC Class II DNA Recombinant Biology Immunoglobulin light chain Molecular biology PBR322 law.invention chemistry.chemical_compound Epitopes Mice Restriction map Plasmid Biochemistry chemistry law Complementary DNA Genetics Recombinant DNA Escherichia coli Methods Animals Immunoglobulin Light Chains Cloning Molecular DNA |
| Zdroj: | Nucleic acids research. 8(9) |
| ISSN: | 0305-1048 |
| Popis: | We have cloned double stranded cDNA sequences encoding a mouse immunoglobulin light chain (L-321) into the PstI site of the beta-lactamase gene of plasmid pBR322 by the oligo (dG)-oligo (dC) tailing procedure. Escherichia coli X1776 transformed by the recombinant plasmids were screened for the expression of L-321 antigenic determinants by a newly developed in situ radio-immunoassay. One out of seven transformants screened was found to synthesize an L-chain like protein. Each bacterial cell produces about 550 molecules of the L-chain sequence. Preferential segregation of the L-chain sequence to the periplasmic space suggest covalent attachment of the L-chain sequence to the N-terminal portion of beta-lactamase. Restriction mapping of the plasmid DNA isolated from the positive clone indicated the presence of a DNA sequence coding for the entire constant region and extending into the variable region for a length corresponding to about 40 amino acid residues. The orientation of the cloned cDNA with respect to the plasmid DNA is compatible with the formation of a fused beta-lactamase-L-321 peptide. |
| Databáze: | OpenAIRE |
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