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1

Application of PCR for the detection of FMD viral RNA

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Paprocka J., Kesy A., Fitzner A., Niedbalski W., Plucienniczak G.
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issue 4
55-61
EN
The polymerase chain reaction (PCR) method has been applied to the detection of FMD viral RNA in samples taken during the clinical stage of FMD from calf. Total RNA was extracted with guanidinum thiocyanate-phenol-chloroform method and reversety transcribed using AMV-reverse transcriptase. cDNA was used as a template for the amplification by PCR of the 672 bp of the VP1 coding sequence.The amplified fragment of cDNA was cloned in the phagemid pBS(+). The first DNA strand was sequenced and consistently an amino acid sequence was established. Comparison between VP1 amino acid sequence of FMDV types C and earlier described type O was performed.
2

Conservation of ferritin amino acid sequences

100%
Smol J.
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issue 4
135-139
EN
The present review describes the conservation of ferritin amino acid sequences. We investigated the N-terminal amino acid sequence of lupin ferritin. We found that both subunits had the same N-terminal sequence. Our data confirm that subunits are not programmed by different mRNAs but they are a result of posttranslational modification. The smaller subunit has a lower molecular weight probably because of deletion of some amino acids from the C-terminal end. Then we compared the lupin ferritin sequence with other known ferritin sequences from plants and animals. Sequence data showed that the identity of plant sequences was 60-92,5%. This high sequence similarity was improved by analysis of genomic DNA from maize and soybean. Both DNAs share 72,2% nucleotide sequence identity. An additional plant specific sequence (named the transit sequence) was observed at the N-terminal end. This sequence consists of 50 nucleotides. We found the lupin ferritin did not have that sequence. We suggest that the deletion of the transit sequence in lupin ferritin seems to be the result of isolation and purification.
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