[Screening and detecting of proteins interaction with KyoT].
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.
Li
Rong
R
Department of Genetics and Developmental Biology, The Fourth Military Medical University, Xi'an 710032, China.
Wang
Ji-Shu
JS
Sun
Qiang
Q
Wang
Jian
J
Yang
Xi
X
Huang
Hong-Yan
HY
Zhou
Peng
P
Han
Hua
H
D003001:Cloning, Molecular / Q000379:methods; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015723:Gene Library; D018398:Homeodomain Proteins / Q000737:chemistry*; D006801:Humans; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D009124:Muscle Proteins; D009687:Nuclear Proteins; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D015003:Yeasts
D004268:DNA-Binding Proteins; C101149:FHL1 protein, human; D018398:Homeodomain Proteins; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D009124:Muscle Proteins; D009687:Nuclear Proteins; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins; C497570:RBPJ protein, human; D014157:Transcription Factors