RT-PCR Analysis of Prn-p mRNA Expression in Normal and Scrapie-Infected B6.1(Prn-pb) . B6(Prn-pa). B6xB6.1(Prn-pa^b) , Tgl5 (Prn-p2a/3b) and Prn-pablated Mice

Abstract

Brains and spleens were extracted from B6.I, B6, FI, Tgl5, and ablated strains of mice, both normal and scrapieinfected. Each strain exhibits a different incubation time due to slight differences found in the allele coding for Prnp mRNA. Total RNA was isolated from these organs. Absorbance readings at 260 nm and 280 nm were used to calculate the concentration of total RNA for each tissue. The reverse transcriptase polymerase chain reaction (RTPCR) was then used to identify different Prn-p alleles and to determine if a quantitative difference in Prn-p mRNA expression exists in the aforementioned mouse strains. Successful distinction and amplification of several Prnp mRNA alleles using RT-PCR has laid the groundwork for subsequent quantitative analyses of Prn-p mRNA.