Selection of HPRT primers as controls for determination of mRNA expression in dogs by RT-PCR
| Autor: | Karen S. Sellins, Glenn R. Frank, Christine M. Johnson, Shumin Yang |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Genetics
Hypoxanthine Phosphoribosyltransferase Base Sequence General Veterinary Reverse Transcriptase Polymerase Chain Reaction Pseudogene Molecular Sequence Data Immunology Sequence Analysis DNA Biology Molecular biology Housekeeping gene genomic DNA Dogs Real-time polymerase chain reaction Complementary DNA Gene expression Animals Amino Acid Sequence RNA Messenger Northern blot Primer (molecular biology) Sequence Alignment Pseudogenes DNA Primers |
| Zdroj: | Veterinary Immunology and Immunopathology. 99:47-51 |
| ISSN: | 0165-2427 |
| DOI: | 10.1016/j.vetimm.2004.01.008 |
| Popis: | Reliable housekeeping gene controls are critical for measuring and comparing gene expression at the transcription level by Northern blot and RT-PCR. In order to develop such controls for studying cytokine mRNA expression in dogs, DNA sequence encoding a full-length canine HPRT protein has been obtained. Numerous primer pairs derived from the canine HPRT sequence have been tested on canine genomic DNA as well as cDNA. The data from the present study suggest that there may be processed HPRT pseudogenes in dogs. Three pairs of canine HPRT primers designed and tested in the present study were able to differentiate between cDNA and genomic DNA under specific PCR conditions. These primers would be useful controls for measurement of mRNA expression by RT-PCR in the dog. |
| Databáze: | OpenAIRE |
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