Construction of a modular yeast two-hybrid cDNA library from human EST clones for the human genome protein linkage map.
Identification of all human protein-protein interactions will lead to a global human protein linkage map that will provide important information for functional genomics studies. The yeast two-hybrid system is a powerful molecular genetic approach for studying protein-protein interactions. To apply this technology to generate a human protein linkage map, the first step is to construct two-hybrid cDNA libraries that cover the entire human genome. With a homologous recombination-mediated approach, we have constructed a modular human EST-derived yeast two-hybrid library in the Gal4 activation domain-based vector, pACT2. Quality analysis of this library indicated that the approach of constructing two-hybrid cDNA libraries from individually arrayed human EST clones is feasible, and such a two-hybrid library is suitable for detecting protein-protein interactions. This is also the first time that a comprehensive two-hybrid system cDNA library has been constructed from a collection of individually arrayed EST clones.
Gene, Net Group, CLONTECH Laboratories Inc., 1020 East Meadow Circle, Palo Alto, CA 94303, USA. sbhua@clontech.com
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SB
Hua
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Luo
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Qiu
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Chan
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Zhou
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Zhu
D002874:Chromosome Mapping; D003001:Cloning, Molecular; D015853:Cysteine Proteinase Inhibitors / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics*; D004274:DNA, Recombinant; D015723:Gene Library; D005800:Genes, Fungal / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017422:Sequence Analysis, DNA; D016324:Sequence Tagged Sites
D015853:Cysteine Proteinase Inhibitors; D018076:DNA, Complementary; D004274:DNA, Recombinant; D011506:Proteins; D011994:Recombinant Proteins