Expression and characterization of a recombinant novel reverse transcriptase of a porcine endogenous retrovirus
| Autor: | Amnon Hizi, Orna Avidan, Ziv Sevilya, Shoshana Loya, Ralf R. Tönjes |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Swine
DNA polymerase Molecular Sequence Data Ribonuclease H Endogenous retrovirus Porcine endogenous virus DNA-Directed DNA Polymerase law.invention Recombinant expression Plasmid law Virology Murine leukemia virus Reverse transcriptase Animals Amino Acid Sequence RNase H Catalytic activities Nucleic Acid Synthesis Inhibitors Manganese biology RNA-Directed DNA Polymerase Endogenous Retroviruses biology.organism_classification Molecular biology Recombinant Proteins Kinetics Protein Subunits Drug resistance DNA Viral biology.protein Recombinant DNA Xenotransplantation |
| Zdroj: | Virology. 307:341-357 |
| ISSN: | 0042-6822 |
| Popis: | The study of porcine endogenous retroviruses (PERVs) becomes increasingly important due to the potential use of pig cells, tissues, and organs as a source for xenogenic cell therapy and xenotransplantation into humans. Consequently, we have constructed a plasmid that induces in bacteria the synthesis of a soluble and highly active reverse transcriptase (RT) of PERV-B. The purified PERV RT was studied biochemically in comparison with the RT of murine leukemia virus (MLV), because of the high-sequence homology between these two RTs. The data show that in several properties the two enzymes are similar, particularly regarding the monomeric subunit composition of the proteins in solution, the high resistance to deoxynucleoside analogues, and the pattern of RNA cleavage by the ribonuclease H activity (RNase H) of the RTs. However, in several cases there are apparent differences between the two RTs, most notable the divalent cation preference (Mn+2 versus Mg+2) in the DNA polymerase reactions. As already shown for viral PERV RT, the novel recombinant PERV RT exhibits a relatively high resistance to several deoxynucleoside analogue inhibitors, suggesting that they might not be very efficient in inhibiting the replication of PERV virions. Therefore, the availability of large amounts of the recombinant RT can be useful for a wide screening of novel drugs against infectious PERV. |
| Databáze: | OpenAIRE |
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