Human Embryonic Hippocampal Yeast Two-hybrid Cdna Library Construction And The Fmr1, Ps1, Ps2 Interacting Proteins

The yeast two-hybrid system is a powerful tool to study protein-protein interaction. Upon the coming of post-genomics era, general yeast two-hybrid strategy is being challenged to adapt library-scale screening proteins interaction. The alternative way reported is using a pool of proteins, or even all predicted ORFs, to screen a library. Such global screening might not only bring breakthrough about in the basic research field, but also commercial profit.Starting from human hippocampus fetus RNA, 2 cDNA libraries were generated, one of which was constructed in DBD vector pGBT9, and the other was constructed in DAD vector pGAD10. The former consisted of 1 × 10~6 cfu and 60% recombinant colonies, the average length of inserts was 1.0 kb; For the latter library, it was made up of 2.5 × 10~6cfu and 90% recombinant colonies, the average length of inserts was 1.5 kb.Among the function study, genes related with neurological diseases attracts many neuroscientists interest. Fragile X mental retardation gene 1(FMR1) is the cause of fragile X syndrome. Presenilin-1(PS1) and Presenilin-2(PS2) is linked with familial Alzheimer's disease. This work is divided into two following parts to study proteins interacting with above three genes.1. Screening of proteins interacted with FMRPFragile X syndrome is the most common cause of inherited mental retardation. Down-regulation of FMR1 results in various clinical manifestations. The role of its protein product FMRP might be a shuttle protein, which transports RNA between cytoplasm and nuclear. The cell process FMRP involvs in still remains understood...