A novel tyrosine kinase, hyk, expressed in murine embryonic stem cells

Horita, K; Yagi, Takeshi; Kohmura, Naohiro; Tomooka, Yasuhiro; Ikawa, Yoji; Aizawa, Shinichi

doi:10.1016/0006-291x(92)90280-x

Cited by 9 publications

(6 citation statements)

References 22 publications

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“…The cDNA library of TT2 ES cells (Horita et al, 1992) was screened with the DNA fragment amplified by 3'RACE in PAT-12. Positive plaques were isolated by the standard method (Sambrook et al, 1989), and the cDNA inserts were subcloned into pBluescriptlI SK(+).…”

Section: Isolation Of Cdna and Sequence Analysismentioning

confidence: 99%

A new strategy of gene trapping in ES cells using 3'RACE

Yoshida

Yagi

Furuta

et al. 1995

Transgenic Research

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"Gene trapping" in embryonic stem (ES) cells is a novel approach to identify a series of genes in mammals concomitant with the production of the corresponding mutant mice. However, this approach is currently unable to identify genes that are not expressed in ES cells. Here we describe a strategy to identify gene trapping clones which is not based on expression of a reporter gene. It uses the neor gene which lacks a polyadenylation signal and has a splice donor signal. Expression of the neor gene as fusion transcripts with the 3' end containing the polyadenylation signal of tagged genes allows the identification of these clones by 3' rapid amplification of the cDNA end in undifferentiated ES cells, even if the genes are not expressed in ES cells. Amplification was observed in about 25% of G418-resistant clones. Sequence analyses suggested the amplifications represent gene trapping events. The feasibility of this approach was further assessed by analysing one clone, PAT-12, in detail.

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Section: Isolation Of Cdna and Sequence Analysismentioning

confidence: 99%

A new strategy of gene trapping in ES cells using 3'RACE

Yoshida

Yagi

Furuta

et al. 1995

Transgenic Research

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“…One matched the sequence of elongation factor-2, and eight matched sequences of the known PTKs FGFR (38), FGFR4 (48), Jakl (75), Jak2 (19), IGFI-R (13), Ryk (26)/Vik (30), Tek (83)/Hyk (25), and Hck (50) (Fig. iB).…”

Section: Resultsmentioning

confidence: 99%

“…The cell suspensions were pooled for every four clones, and the cell pellets were resuspended in 50 ,ul of water, boiled for 10 min, treated with 5 ,ug of proteinase K at 55°C for 120 min, and boiled again for 10 min. Half of the mixture was used as a template for PCR analyses; the analyses were made with 25 pmol each of two primers, 5'-TTGACGAGTTCTITCTGA-3' and 5'-GACCAG GACCAGTCTGAACACATCCATCCG-3'; the former is located at the 3' terminus of the neor gene, and latter is located at the 3' end outside of the targeting vector in the srm gene (see Fig. 5A).…”

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confidence: 99%

A novel nonreceptor tyrosine kinase, Srm: cloning and targeted disruption.

Kohmura¹,

Yagi²,

Tomooka³

et al. 1994

Mol. Cell. Biol.

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“…Wie bei VEGF entsteht die selektive Wirkung der Angiopoietine am Gefäßendothel durch den spezifischen Rezeptorbesatz. Die Rezeptoren werden als Tie1 und Tie2 bezeichnet und sind Tyrosinkinasen [2,6,7,13].…”

Section: Angiopoietineunclassified