Method for cloning genes
US4394443A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Dec 18, 1980 |
| Grant date | Jul 19, 1983 |
| Priority date | — |
| Expiry date | Dec 18, 2000 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1096
- WIPO fieldOrganic fine chemistry
- WIPO sectorChemistry
Abstract
A method is provided for isolating and identifying a recombinant clone having a DNA segment therein coding for at least one desired heterologous polypeptide, at least a short amino acid sequence of which is known, by effecting cDNA synthesis on a mixture of mRNAs containing the mRNA coding for the desired polypeptide, isolating the resultant cDNA mixture, inserting the resultant cDNA into recombinant cloning vehicles, transforming hosts with the vehicles, separating the transformants and isolating and identifying a recombinant clone containing a DNA segment which is homologous over at least a portion thereof to at least one oligonucleotide probe specific for the DNA segment; wherein the probe is an extension of the nucleotide sequence of an oligonucleotide primer having a nucleotide sequence complementary to a region of the target mRNA coding for a portion of the known amino acid sequence, and is complementary to a longer region of the target mRNA coding for a longer portion of the known amino acid sequence. Recombinant clones coding for human histocompatibility antigens are provided, in particular, clones for HLA-B antigens.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.