REAGENTS/MATERIAL: Human Bcl-2 cNDA, kind gift from Dr. Walter Nishioka (Vical Inc. San Diego, CA)
PrP and PrP-D178N and -T183A DNAs were PCR-amplified.
Bax-a cDNA was PCR-amplified from human neuron cNDA.
cDNAs were cloned into pBluescript KSII (pBSK-; Stratagene) or pCep4b (Invitrogen)
Deletion of OR region by PCR amplification with the following primers:
Human primary neurons were cultured and microinjected with 25 pl containing 0.75 pg of DNA and 2.5 pg of dextran Texas Red in phosphate buffered saline.
FUTURE DIRECTION: Further studies needed to understand the underlying molecular mechanism of PrP function against Bax. Also, to determined whether FFI and GASE PrP mutation undergo a loss of function against Bax-mediated cell death is due to altered protein conformation or improper trafficking.