Construction of a modular yeast two-hybrid cDNA library from human EST clones for the human genome protein linkage map
1998; Elsevier BV; Volume: 215; Issue: 1 Linguagem: Inglês
10.1016/s0378-1119(98)00258-3
ISSN1879-0038
AutoresShao-bing Hua, Ying Luo, Mengsheng Qiu, Eva Chan, Helen Zhou, Li Zhu,
Tópico(s)RNA Research and Splicing
ResumoIdentification 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.
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