Rapid isolation of cell-type-specific protein tyrosine kinases by degenerate polymerase chain reaction combined with differential hybridization technique
| Autor: | Tadao Kakizoe, Masaru Katoh, Atsushi Takahashi, Sun Jin Kim, Masaaki Terada, Takashi Sugimura, Hiroshi Sasaki, Y.K. Kim |
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| Rok vydání: | 1995 |
| Předmět: |
animal structures
DNA Complementary Molecular Sequence Data Biophysics Biology Biochemistry Polymerase Chain Reaction law.invention law Renal cell carcinoma Complementary DNA Proto-Oncogene Proteins medicine Tumor Cells Cultured Molecular Biology Gene Polymerase chain reaction Base Sequence RNA Cancer Nucleic Acid Hybridization Receptor Protein-Tyrosine Kinases Cell Biology Protein-Tyrosine Kinases medicine.disease Molecular biology fms-Like Tyrosine Kinase 3 Cell culture Tyrosine kinase |
| Zdroj: | Biochemical and biophysical research communications. 214(1) |
| ISSN: | 0006-291X |
| Popis: | To identify protein tyrosine kinase (PTK) genes preferentially expressed in renal cell carcinoma cell line, we screened a PTK-cDNA-enriched library constructed from RNA of an renal cell carcinoma cell line with a PTK probe, each produced from renal cell carcinoma, gastric cancer or esophageal cancer cell lines by degenerate polymerase chain reaction. Two cDNA fragments of PTK genes, FRK and FLT-3, were isolated from the PTK-cDNA-enriched library of the renal cell carcinoma cell line by differential hybridization technique. The FRK cDNA clone represented 15.8% of the PTK-cDNA-enriched library from the renal cell carcinoma cell line, while the FLT-3 cDNA clone was 2.8% of the same library. Both of the two PTK genes were expressed preferentially in renal cell carcinoma cell lines. This method, described here, is useful for the rapid isolation of PTK cDNA fragments, including a low abundant cDNA, preferentially expressed in a specific cell line. |
| Databáze: | OpenAIRE |
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