Cloning of JNK activation domain and Fas-binding domain of human apoptotic Daxx gene by homologous recombination in yeast
DOI:
https://doi.org/10.14456/tjg.2008.11Keywords:
cloning, human Daxx, JNK activation domain, Fas-binding domain, yeastAbstract
The death domain-associated protein (Daxx) was originally cloned as a Fas-interacting protein. It is ubiquitously expressed and highly conserved in mammals. Currently, the role of Daxx is controversy between apoptotic or anti-apoptotic molecules. To understand the precise function of human Daxx, yeast two hybrid screening will be used as a tool to identify interactions between human Daxx and other human proteins. The first step in a yeast two hybrid method is to express a bait plasmid in the recombinant form in yeast. We utilized the in vivo homologous recombination in yeast to insert a JNK activation domain and a Fas-binding domain of human Daxx, into the bait plasmid, pEG-NRT. Furthermore, the expression of a JNK activation domain and a Fas-binding domain of human Daxx was demonstrated by Western blot analysis. Yeast expressing a JNK activation domain and a Fas-binding domain of human Daxx will be used as bait strains to screen for human proteins interacting with Daxx, which may provide us the clue to uncover the mysterious function of human Daxx.
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