RRC ID 1241
著者 Sasaki E, Hikono H, Kaku Y, Kuwana T, Naito M, Sakurai M.
タイトル ephA9, a novel avian receptor tyrosine kinase gene.
ジャーナル Gene
Abstract Protein tyrosine kinase (TK) receptors are known to play crucial roles in various aspects of normal development and in tumorigenesis. Germ cells before their colonizing to the gonads during embryogenesis are called primordial germ cells (PGCs). To identify TK genes expressed in chicken PGCs, we purified these cells from the blood of 2.5-day-old embryos, extracted the polyA(+) RNA, and subjected it to reverse transcription-polymerase chain reaction (RT-PCR) amplification with TK gene-specific primers. As a result, we identified 13 receptor TK genes and 6 non-receptor TK genes. One of the receptor TK genes appeared to be novel, and thus the full-length DNA complementary to RNA (cDNA) sequence was retrieved by the rapid cloning of cDNA ends method. Sequence analysis of this cDNA indicated that it encoded a novel TK receptor of the Eph family. The putative amino-acid sequence of this TK was 63.0% identical to that of human EphA1; therefore, we designated our novel TK as EphA9. Northern blot hybridizations indicated that ephA9 RNA transcripts were present in the kidney, lung, testis, and thymus but not in the spleen, brain, or liver. Expression of a fusion protein containing the intracellular domain of EphA9 in bacterial cells showed that this domain had TK enzymatic activity. The EphA9 species produced in Cos-1 cells transfected with an expression vector were tyrosine-phosphorylated. These data suggest that EphA9 plays its biological role(s) in various organs of chickens as an active TK.
巻・号 316
ページ 103-10
公開日 2003-10-16
DOI 10.1016/s0378-1119(03)00743-1
PII S0378111903007431
PMID 14563556
MeSH Amino Acid Sequence Animals Blotting, Northern COS Cells Chick Embryo Chickens / genetics* Chlorocebus aethiops DNA, Complementary / chemistry DNA, Complementary / genetics Gene Expression Regulation, Enzymologic Glutathione Transferase / genetics Glutathione Transferase / metabolism Male Molecular Sequence Data Polymerase Chain Reaction RNA, Messenger / genetics RNA, Messenger / metabolism Receptor Protein-Tyrosine Kinases / genetics* Receptor Protein-Tyrosine Kinases / metabolism Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Sequence Analysis, DNA Sequence Homology, Amino Acid Transfection
IF 2.984
引用数 6
WOS 分野 GENETICS & HEREDITY
リソース情報
ヒト・動物細胞 COS-1(RCB0143)