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Table 1 Summary of characterization of properties of EK211 cattle

From: Effects of a naturally occurring amino acid substitution in bovine PrP: a model for inherited prion disease in a natural host species

1

2

3

4

5

6

7

8

Calf number

Genotype

% WT Allele (G)/% K Allele (A)

Ct

(PRNP)

Ct

(ACTB)

ΔCt

Plasma TBARS: OD540 reading

Plasma SOD level (U/ml)

#79

EE211

–

30.4

19.6

10.8

0.18

0.036

#84

EK211

54/46

30.0

19.0

11.0

0.19

0.040

#85

EE211

–

30.0

19.2

10.8

0.15

0.046

#86

EE211

–

30.9

20.3

10.6

0.20

0.046

#87

EK211

54/46

30.0

18.9

11.2

0.20

0.055

#88

EK211

52/48

30.6

18.9

11.6

0.17

0.054

EE211 Average

 

10.8 ± 0.1

0.18 ± 0.03

0.043 ± 0.007

EK211 Average

 

11.3 ± 0.3

0.19 ± 0.02

0.052 ± 0.008

  1. Column 3 presents the results of allele expression analysis of RNA from EK211 cattle PBLs. Ratios were assessed by analysis of sequencing reactions of PCR amplicons of the region around codon 211 in PRNP cDNA, using QSVAnalyzer as described in additional methods (Additional file 1). Cattle were 48 months of age at the time of this analysis. Columns 4–6 represent the results of qRT-PCR analysis of PRNP expression levels in both EE211 and EK211 cattle PBLs, with Column 6 indicating the ΔCt = Ct(PRNP) − Ct(ACTB). Column 7 depicts results of TBARS readings on peripheral blood plasma samples. TBARS numbers reflect the OD540 of the cattle plasma samples as measured by the TBARS Kit from Cayman Chemical; due to interference from a separate absorbance peak in these samples, values are reported here as absorbance at 540 nm as opposed to a quantitative value based on the standard curve. Column 8 depicts the results of SOD (Superoxide Dismutase) assays on peripheral blood plasma samples. SOD values are expressed in U/ml as derived from a standard curve generated with the SOD Assay Kit from Cayman Chemical. For columns 4–8, animals were 26 months of age at the time of analysis, with the exception of animal #79, which was 32 months at the time. The bottom two rows depict average values across biological replicates for parameters, expressed ± the 95% CI (confidence interval) in each case. (We note that we assume a normal distribution to calculate the CI; due to the unique nature of this cattle population, a small number of EK211 animals are available for testing, precluding more in-depth examinations of normality). The SOD assay on peripheral blood plasma was also completed on the offspring of the U.S. 2006 H-type BSE case (level = 0.057), and this value was included in the EK211 cattle average (Column 8)
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BMC Research Notes

ISSN: 1756-0500

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