PCR mediated detection of a new human receptor-tyrosine-kinase, HEK 2
| Autor: | Böhme B, Holtrich U, Wolf G, Luzius H, Kh, Grzeschik, Klaus Strebhardt, Rübsamen-Waigmann H |
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| Rok vydání: | 1993 |
| Předmět: |
Amyloid
Base Sequence Receptor EphA2 Receptor EphB3 Molecular Sequence Data Receptor EphA3 Membrane Proteins Receptors Cell Surface Protein-Tyrosine Kinases Polymerase Chain Reaction Cell Line Neoplasm Proteins Rats Blotting Southern Open Reading Frames Animals Humans Amino Acid Sequence Conserved Sequence |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0950-9232 |
| Popis: | We have previously amplified cDNA subfragments of protein-tyrosine-kinases (PTKs) by using the polymerase chain reaction (PCR) and specific sets of oligonucleotide primers derived from nucleotide sequences of their kinase domain. In this study we have used a more directed approach to identify new members of the EPH/elk-family by PCR of human embryonic cDNA: we utilized oligonucleotide primers specifically designed to a highly conserved N-terminal motif and the kinase region of EPH/elk-PTKs in RNA-PCRs. The 5' and 3' elongation of the primary PCR product was achieved by the RACE (rapid amplification of cDNA ends)-technique. Sequence analysis of 3.8 kb of overlapping PCR products allowed to identify a novel receptor-PTK, HEK 2 (human embryo kinase 2), as an additional member of this family, without the need to screen a cDNA library. This approach should be useful for the rapid isolation of other PTK-genes as well. Analysis of genomic DNA placed HEK 2 on chromosome 3. Northern blot analysis demonstrated the expression of a 4.6 kb HEK 2-mRNA in lung, brain, pancreas, liver, placenta, kidney, skeletal muscle, heart and several human cells. In a protein kinase assay with HEK 2-specific immunoprecipitates from the human epidermoid carcinoma cell line A431, a protein of 130 kDa was found phosphorylated. |
| Databáze: | OpenAIRE |
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