Rapid isolation of cell-type-specific protein tyrosine kinases by degenerate polymerase chain reaction combined with differential hybridization technique.
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.
Kim
S J
SJ
Genetics Division, National Cancer Center Research Institute, Tokyo, Japan.
Sasaki
H
H
Takahashi
A
A
Katoh
M
M
Kakizoe
T
T
Kim
Y K
YK
Sugimura
T
T
Terada
M
M
D001483:Base Sequence; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000302:isolation & purification*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000302:isolation & purification*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000302:isolation & purification*; D014407:Tumor Cells, Cultured; D051941:fms-Like Tyrosine Kinase 3
D018076:DNA, Complementary; D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3