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

Research Project

Project/Area Number 09470086
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Virology
Research InstitutionNAGASAKI 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
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsprion / 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.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report

URL: 

Published: 1998-04-01   Modified: 2016-04-21  

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