Ababhali: Wang Xiaoyan, Zhao Eryu

Iyunithi: Isibhedlela i-Jiaozhou, Isibhedlela sase-Dongfang Esixhumene neNyuvesi yase-Tongji

Njengamanje, uhlobo lwesampula oluyinhloko lokutholwa kwe-pathogen nucleic acid yokuphefumula ukuswabha komphimbo.Kunezinhlobo eziningi zezixazululo zokulondoloza isampula ezivame ukusetshenziswa, zonke ezidinga ukufakwa esiqandisini noma ziqandiswe ukuze zihanjiswe futhi zigcinwe;Kunzima ukulawula yonke inqubo yokushisa okuphansi ngesikhathi sokuqoqwa kanye nenqubo yokuthutha, futhi kunzima ukuqinisekisa ukulawulwa kwekhwalithi ngaphambi kokuhlolwa kwesampula.^[1-2].

I-RNase (RNase) iyi-endonuclease eyenza i-RNA i-hydrolyze, ikakhulukazi inqamula izibopho ze-phosphodiester phakathi kwama-nucleotide.I-molecule ye-RNase izinzile kakhulu, kunezibopho ze-disulfide esakhiweni, futhi umsebenzi wayo awudingi ukuba khona kwama-divalent cations, ngakho-ke i-RNase ayishintshiwe kalula, futhi kulula ukuyibuyisela kabusha ngisho nangemva kokushisa okuphezulu noma ukusetshenziswa kwama-denaturants.Ama-RNase ahlukaniswe abe yi-endogenous kanye ne-exogenous.Ama-RNases angapheli angakhululwa ngesikhathi esifanayo lapho amaseli ephukile.Ngakho-ke, ukuqeda indima yama-RNases angapheli kuyisinyathelo esibucayi kakhulu kunqubo yokukhipha i-RNA.Ama-RNase angaphandle asatshalaliswa kabanzi.Ama-RNase akhona emoyeni, esikhumbeni somuntu, izinwele namathe, okuyizizathu ezibalulekile zokuwohloka kalula kwe-RNA.^[3].

Umbuzo wedatha

I-CANS-CL02-A009 “Ukusetshenziswa Kwemihlahlandlela Yekhwalithi Yelabhorethri Yezokwelapha Nokufaneleka Kokugunyazwa Emkhakheni Wokuxilonga Amangqamuzana” ezimfuneko zobuchwepheshe iphakamisa ukuthi amazinga afanelekile ekhwalithi yamanzi kufanele akhiwe ngokuvumelana nesicelo;Iziqukathi ezingangeni moya ezilahlwayo:

I-RNase/Ukuhlukaniswa

(1) RNase A

I-Ribonuclease A (RNase A), etholakala ku-pancreas yezinkomo, iyi-endoribonuclease engahlasela ngokuqondile ukuphela kwe-3' yezinsalela ze-pyrimidine ku-RNA, ukusika i-cytosine noma i-uracil eyakhiwe ama-nucleotide aseduze.Isibopho se-Phosphodiester, umkhiqizo wokugcina wokusabela yi-3 'pyrimidine nucleotide kanye ne-oligonucleotide ene-3' pyrimidine nucleotide ekugcineni.

(2) I-RNase T1

I-Ribonuclease T1 (RNase T1) isuselwa ku-Aspergillus orjzae, isebenza ngokukhethekile ku-3′-terminal phosphate ye-guanine, futhi indawo yokuqhekeka iphakathi kwe-3′ phosphate ye-guanine kanye ne-5′ hydroxyl yama-nucleotide aseduze.Umkhiqizo wokugcina wokusabela yi-3′guanylic acid nezingcezu ze-oligonucleotide ezine-3′guanylic acid ekugcineni.

(3) RNase H

I-Ribonuclease H (RNase H) yatholwa okokuqala esiculweni sethole le-thymus, futhi isakhi sayo sombhalo wekhodi senziwe saba yi-Escherichia coli.Ingalulaza ngokuqondile i-DNA: Imicu ye-RNA kuma-RNA hybrid duplexes, okuholela kuma-oligonucleotides nama-mononucleotide ane-3′-OH kanye no-5′-monophosphate ekupheleni, ayikwazi ukwehlisa i-DNA eyodwa noma kabili enemicu emibili noma i-RNA.

RNase

Umsebenzi kanye nokusetshenziswa

I-Ribonuclease ingadala ukuwohloka kwe-ribonucleic acid (RNA) futhi ingahlanganiswa ngokwenziwa.Amafutha okwelapha asetshenziselwa ukwelapha ukuhlukumezeka nobuhlungu bamalunga.Ngokwemibiko, i-ribonuclease ingashintsha i-host cell metabolism, ivimbele ukuhlanganiswa kwegciwane, ivimbele ukwanda kwegciwane lomkhuhlane ku-vitro, futhi ivimbele ukwakheka kwegciwane le-vaccinia ne-herpes emibungu yenkukhu.Ukusetshenziswa komtholampilo komjovo we-ribonuclease nsuku zonke we-intramuscular of 180 mg, okuzuzisayo ekwelapheni ubhubhane lwe-encephalitis, i-ribonuclease ingadala ukuwohloka kwe-ribonucleic acid (RNA), futhi manje isingenziwa ngokwenziwa.Amafutha okwelapha asetshenziselwa ukwelapha ukuhlukumezeka nobuhlungu bamalunga.Ngokwemibiko, i-ribonuclease ingashintsha i-host cell metabolism, ivimbele ukuhlanganiswa kwegciwane, ivimbele ukwanda kwegciwane lomkhuhlane ku-vitro, futhi ivimbele ukwakheka kwegciwane le-vaccinia ne-herpes emibungu yenkukhu.Ukusetshenziswa komtholampilo komjovo we-ribonuclease nsuku zonke we-180 mg kunenzuzo ekwelapheni ubhubhane lwe-encephalitis.

RNase

Inhibitor Definition

Incazelo yeyunithi ye-RNase inhibitor: Inani le-enzyme edingekayo ukuze kuvinjwe u-50% womsebenzi we-5ng RNase A iyunithi eyodwa.

Isebenza kanjani i-RNase Inhibitors

I-Guanidine isothiocyanate:

I-Guanidine isothiocyanate iyinhlanganisela ye-organic ene-molecular formula C2H6N4S.Ikakhulukazi esetshenziswa emithini yezinto eziphilayo, ama-reagents amakhemikhali, njll. I-Guanidine isothiocyanate iyi-protein lysing agent, futhi ivame ukusetshenziswa njengengxenye eyinhloko yesisombululo se-lysis kuma-reagents okuxilongwa kwamangqamuzana.Ingakwazi ukuhlanganisa izicubu, ibhubhise ukwakheka kweseli futhi ihlukanise i-nucleic acid ku-nucleoprotein, futhi ibe ne-denaturation eqinile ku-RNase.Iyi-RNase inhibitor esebenza kahle kakhulu njengamanje.

I-TRIzol iyinoveli isiyonke ye-RNA extraction reagent engakhipha ngokuqondile i-RNA ephelele kumaseli noma izicubu.Iqukethe izinto ezifana ne-phenol ne-guanidine isothiocyanate, ezingaphazamisa ngokushesha amangqamuzana futhi zivimbele ama-nuclease akhishwa amaseli.

(Kodwa-ke, i-guanidine isothiocyanate iyingozi empilweni yabacwaningi belebhu.)

I-RNasin:

I-Acid glycoprotein ekhishwa esibindini segundane noma ku-blastoderm yomuntu.I-Rnasin iyi-inhibitor engaqhudelani ye-RNase, engabophezela kuma-RNa ahlukahlukene ukuze iwavule.

(Imininingwane eyengeziwe: https://www.foreivd.com/foreasy-rnase-inhibitor-product/)

Hizinga lokushisa le-igh:

Ukushisa okuphezulu futhi kuyindlela evamile yokuguqulwa kwamaprotheni.

I-Diethylpyrocarbonate (DEPC):

I-DEPC iyisivimbeli se-RNase esiqinile kodwa esingaphelele, esingavimbela umsebenzi we-RNase ngokuhlanganisa iringi ye-amino acid imidazole yeqembu elisebenzayo le-RNase ukuze iguqule amaprotheni.

I-Vanadyl ribonucleoside complex:

Inkimbinkimbi eyakhiwe i-vanadium oxide ion nama-nucleosides, ebophezela ku-RNase ngendlela yezinto eziguquguqukayo, engacishe ivimbe ngokuphelele umsebenzi we-RNase.

okunye:

I-SDS, i-urea, umhlaba we-diatomaceous, njll. nakho kunomphumela othile ovimbelayo ku-RNase.

Izibuyekezo Zochwepheshe

U-Li Yujie Uchwepheshe Oyinhloko

Umqondisi woMnyango Welebhu, Isibhedlela sase-Jiaozhou, Isibhedlela sase-Dongfang Esixhumene ne-Tongji University

Ukuze uvimbele ukubola kwe-RNase yangaphandle, imaski, amagilavu ​​nezigqoko kufanele kugqokwe futhi kushintshwe njalo phakathi nenqubo yokukhipha i-RNA.Zonke izitsha zengilazi kufanele zibhakwe kuhhavini owomisa ku-200 ° C amahora angaphezu kwama-2.Izinto ezisetshenziselwa ukubhaka, njengepulasitiki, zidinga ukuphathwa ngamanzi e-DEPC, bese ziwashwa ngamanzi acwecwe.Ama-reagents noma izisetshenziswa ezisetshenziselwa ukukhipha, ukugcinwa kanye nokuhlonza i-RNA kufanele zinikezelwe ku-RNA, futhi kumiswe indawo yokusebenza ye-RNA ezimele.

izinkomba:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.Amagciwane namagciwane ku-nasopharynx ngokushesha ngaphambi kokuqala kwezifo eziyingozi zokuphefumula eziphansi ezinganeni zomdabu wase-Australia[J].I-Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Igatsha Lokulawula Ukutheleleka Esibhedlela le-Chinese Preventive Medicine Association.Imihlahlandlela yokuqoqwa nokuhlolwa kwezibonelo ze-microbial clinic [J].Ijenali yesiShayina Yokutheleleka Kwesibhedlela, 2018(20):3192-3200.

[3] "Ukuhlolwa Komtholampilo Izinkulungwane Eziyishumi Kungani Ivolumu Yokuhlola I-Molecular Biology"