Elucidation of the nature of a Creutfeldt-Jakob disease agent using prion protein gene-knockout mice

1998 Fiscal Year Final Research Report Summary

• Project/Area Number: 09470086
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: NAGASAKI UNIVERSITY
• Principal Investigator: SAKAGUCHI Suehiro Nagasaki University School of Medicine Assistan, 医学部, 助手 (60274635)
• Co-Investigator(Kenkyū-buntansha): SHIGEMATU Kazuto Nagasaki University, School of Medicine Associate Professor, 医学部, 助教授 (20154205)
• Project Period (FY): 1997 – 1998
• Keywords: prion / knockout mouse / liposome

Research Abstract

Prion protein gene-knockout (PrP-/-) mice : Heterozygous (PrP+1-) mice were mated to produce PrP-/- mice, which resulted in 27 PrP-/- mice.
Production of anti-PrP serum : Mouse PrP was expressed in E.coli and purified using ion-exchange chromatography and gel filtration. Anti-PrP sera were taken form PrP-/- mice after immunization with the recombinant mouse PrP.These sera showed nice signals of PrP on western blotting.
Generation of prion liposomes : At first, we elucidated the method for making liposomes using beta-galactosidase DNA.The beta-galactosidase DNA was mixed with phosphatidylcholine and detergent and inoculated into C57BL/6 mice intracerebrally. Their brains were removed 3 days after inoculation and stained by x-gal. We could not detect the xi-galactosidase-positive neurons in brains of 20 mice, indicating clearly that this method we used here was not efficient for introducing of exogenous substances into brain. Although the fraction enriched for prion was prepared from the infected-C57BT/6 mouse brains using differential centrifugation, we could not proceed to make prion liposomes and to inoculation experiment of prion liposomes.