Ishicilelwe ngu: I-CAS Key Laboratory ye-Pathogen Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences
IF: 69.504
Umkhiqizo Osizwayo:Ikhithi Yokuzihlukanisa Yeseli Ye-RNA
Isixhumanisi sangempela:https://www.nature.com/articles/s41586-022-05271-2
Ukutheleleka ngama-filovirus afana ne-Ebola virus (EBOV) kanye ne-Marburg virus kungabangela izimpawu ezinzima zomtholampilo, kuhlanganise ne-hemorrhagic fever kanye nokwehluleka kwezitho eziningi.4.Lapha sinqume ukwakheka kwe-Ebola virus L protein eyinkimbinkimbi ene-tetrameric VP35 kusetshenziswa i-cryo-electron microscopy (isifunda 1).Ukuhlaziywa kwesakhiwo kwembula ukuthi igciwane le-Ebola L linesici sokufaka esikhethekile se-filovirus esibalulekile ekuhlanganiseni kwe-RNA, nokuthi i-VP35 isebenzisana kakhulu nesifunda se-N-terminal sika-L ngama-protomer amathathu e-VP35 tetramer.
Kuleli phepha, i-RNA isiyonke yakhishwa kusetshenziswa i-ForegeneIkhithi Yokuzihlukanisa Yeseli Ye-RNAfuthi ukugxiliswa kwe-RNA kukalwa nge-Nanodrop ku-260 nm, bese yenza ukutholwa kwe-RT/qPCR.
■ Yonke inqubo iqhutshwa ekamelweni lokushisa (15-25 ℃), ngaphandle kokugeza kweqhwa kanye nokushisa okuphansi kwe-centrifugation.
■ Ikhithi yonke ayinayo i-RNase-Free, asikho isidingo sokukhathazeka ngokuwohloka kwe-RNA.
■ Ikholomu Yokuhlanza I-DNA ibopha ngokukhethekile i-DNA, ukuze ikhithi ikwazi ukususa ukungcola kwe-DNA ye-genomic ngaphandle kokwengeza i-DNase eyengeziwe.
■ Isivuno esikhulu se-RNA: Ikholomu ye-RNA kuphela kanye nefomula eyingqayizivele ingahlanza i-RNA ngokuphumelelayo.
■ Isivinini esisheshayo: kulula ukusebenza futhi singaqedwa ngemizuzu eyi-11.
■ Ukuphepha: Asikho i-reagent ephilayo edingekayo.
■ Ikhwalithi ephezulu: I-RNA ehlanziwe imsulwa kakhulu, ayinamaprotheni nokunye ukungcola, futhi ingahlangabezana nokuhlola okuhlukahlukene okulandelayo.
Isikhathi sokuthumela: Nov-04-2022
