Sequences within Both the 5′ UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus (MMTV) Genomic RNA 英文参考文献.docVIP

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Sequences within Both the 5′ UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus (MMTV) Genomic RNA 英文参考文献.doc

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Sequences within Both the 5′ UTR and Gag Are Required for Optimal In Vivo Packaging and Propagation of Mouse Mammary Tumor Virus (MMTV) Genomic RNA 英文参考文献

SequenceswithinBoththe59UTRandGagAreRequired forOptimalInVivoPackagingandPropagationofMouse MammaryTumorVirus(MMTV)GenomicRNA FarahMustafa1*,DhuhaAlAmri2,FarahAlAli2,NoorAlSari2,SarahAlSuwaidi2,PreethiJayanth2¤, PrettyS.Philips2,TahirA.Rizvi2 1DepartmentsofBiochemistry,FacultyofMedicineandHealthSciences(FMHS),UnitedArabEmiratesUniversity(UAEU),AlAin,UnitedArabEmirates,2Microbiology Immunology,FacultyofMedicineandHealthSciences(FMHS),UnitedArabEmiratesUniversity(UAEU),AlAin,UnitedArabEmirates Abstract Background: This study mapped regions of genomic RNA (gRNA) important for packaging and propagation of mouse mammarytumorvirus(MMTV).MMTVisatypeBbetaretroviruswhichpreassemblesintracellularly,aphenomenondistinct fromretrovirusesthatassembletheprogenyvirionatcellsurfacejustbeforebuddingsuchasthetypeChumanandfeline immunodeficiencyviruses(HIVandFIV).StudiesofFIVandMason-Pfizermonkeyvirus(MPMV),atypeDbetaretroviruswith similarintracellularvirionassemblyprocessesasMMTV,haveshownthatthe59untranslatedregion(59UTR)and59endof gagconstituteimportantpackagingdeterminantsforgRNA. Methodology: Three series of MMTV transfer vectors containing incremental amounts of gag or 59 UTR sequences, or incrementalamountsof59UTRinthepresenceof400nucleotides(nt)ofgagwereconstructedtodelineatetheextentof59 sequences that may be involved in MMTV gRNA packaging. Real time PCR measured the packaging efficiency of these vector RNAs into MMTV particles generated by co-transfection of MMTV Gag/Pol, vesicular stomatitis virus envelope glycoprotein (VSV-G Env), and individual transfer vectors into human 293T cells. Transfer vector RNA propagation was monitoredbymeasuringtransductionoftargetHeLaT4cellsfollowinginfectionwithviralparticlescontainingahygromycin resistancegeneexpressioncassetteonthepackagedRNA. PrincipalFindings:MMTVrequirestheentire59UTRandaminimumof,120nucleotide(nt)atthe59endofgagfornot onlyefficientgRNApackagingbutalsopropagationofMMTV-basedtransfervectorRNAs.VectorRNAswithouttheentire