Abstract
To study the function of KyoT2 in vivo, two-hybrid yeast, purification of KyoT2 protein, preparation of antibody and GST-pulldown methods were used in the experiments. 42 clones were obtained after 5 x 10(6) clones were screened by four kinds of nutrition limitation and beta-galactosidase assay, 22 clones were obtained after restriction of positive clones. Finally, 13 genes were obtained by sequence assay. Two of these were RBP-Jk and PIAS1. After they and KyoT2 changed vectors, negative two-hybrid yeast was finished. The result was positive; Using KyoT2 protein and antibody GST-pulldown of KyoT2 and RBP-Jk, KyoT2 and PIAS1 were done, the result was also positive. Therefore, KyoT2 interacted with RBP-Jk and PIAS1.
MeSH terms
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Cloning, Molecular / methods
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Gene Library
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Homeodomain Proteins / chemistry*
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Humans
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Immunoglobulin J Recombination Signal Sequence-Binding Protein
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Intracellular Signaling Peptides and Proteins
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LIM Domain Proteins
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Muscle Proteins*
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Nuclear Proteins*
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Protein Inhibitors of Activated STAT
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Proteins / genetics
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Proteins / metabolism*
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Transcription Factors / chemistry
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Transcription Factors / genetics
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Transcription Factors / metabolism*
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Two-Hybrid System Techniques*
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Yeasts
Substances
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DNA-Binding Proteins
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FHL1 protein, human
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Homeodomain Proteins
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Immunoglobulin J Recombination Signal Sequence-Binding Protein
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Intracellular Signaling Peptides and Proteins
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LIM Domain Proteins
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Muscle Proteins
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Nuclear Proteins
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Protein Inhibitors of Activated STAT
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Proteins
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RBPJ protein, human
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Transcription Factors