Tusitala: Wang Xiaoyan, Zhao Eryu

Vaega: Falemai Jiaozhou, Falemai o Dongfang Faʻatasi i le Iunivesite o Tongji

I le taimi nei, o le ituaiga faʻataʻitaʻiga autu mo le suʻesuʻeina o le faʻamaʻi faʻamaʻi faʻamaʻi nucleic acid o le faʻaʻi swab.E tele ituaiga o vaifofo fa'asao fa'ata'ita'iga e masani ona fa'aaogaina, o ia mea uma e mana'omia ona fa'a'aisa pe fa'aisa mo felauaiga ma teuina;E faigata ona pulea le faagasologa atoa o le maualalo o le vevela i le taimi o le aoina ma le felauaiga, ma e faigata ona faʻamautinoa le lelei o le pulea aʻo leʻi faia suʻega faʻataʻitaʻiga.^[1-2].

RNase (RNase) ose endonuclease e fa'asusu ai le RNA, aemaise lava le motusia o sootaga phosphodiester i le va o nucleotides.O le RNase mole e matua mausali, o loʻo i ai faʻamaufaʻailoga disulfide i totonu o le fausaga, ma o lana gaioiga e le manaʻomia ai le i ai o cation divalent, o lea e le faigofie ai le RNase denatured, ma e faigofie ona toe faʻafouina e tusa lava pe maualuga le vevela poʻo le faʻaaogaina o denaturants.RNases ua vaevaeina i totonu ma fafo.E mafai ona fa'asa'oloto RNases i le taimi e tasi pe a malepelepe sela.O le mea lea, o le faʻaumatiaina o le matafaioi a RNases faʻamaʻi o se laasaga sili ona taua i le faʻagasologa o le RNA.Exogenous RNases e salalau lautele.RNases o loʻo i ai i le ea, paʻu o le tagata, lauulu ma faua, o mafuaʻaga taua ia mo le faʻaleagaina faigofie o le RNA.^[3].

Su'ega fa'amatalaga

CANS-CL02-A009 "Faʻaaogaina o Faʻatonuga Faʻapitoa a Fale Suʻesuʻe Faʻafomaʻi ma Taʻiala Faʻamaonia Agavaʻa i le Faʻalapotopotoga o Suʻesuʻega Molecular" i manaʻoga faʻapitoa e faʻatatau e tatau ona faʻatulagaina tulaga lelei o le vai e tusa ai ma le talosaga;Koneteina mālūlū fa'aaoga:

RNase/Fa'avasegaga

(1) RNase A

Ribonuclease A (RNase A), e maua mai le bovine pancreas, o se endoribonuclease e mafai ona osofaʻia faʻapitoa le 3' pito o totoe o pyrimidine i luga o le RNA, tipiina o le cytosine poʻo le uracil e fausia e nucleotides lata ane.Phosphodiester bond, o le mea mulimuli o le tali o se 3' pyrimidine nucleotide ma se oligonucleotide ma se 3' pyrimidine nucleotide i le pito.

(2) RNase T1

Ribonuclease T1 (RNase T1) e maua mai Aspergillus orjzae, e galue faapitoa i luga o le 3'-terminal phosphate o le guanine, ma o le nofoaga cleavage i le va o le 3' phosphate o le guanine ma le 5' hydroxyl o nucleotides lata ane.O le mea mulimuli o le tali o le 3'guanylic acid ma vaega oligonucleotide ma le 3'guanylic acid i le pito.

(3) RNase H

O le Ribonuclease H (RNase H) na maua muamua mai le aano o le tama'i povi, ma o lona fa'asologa o kene na fa'apipi'iina i le Escherichia coli.E mafai ona faʻaleagaina faʻapitoa DNA: RNA strands i RNA hybrid duplexes, e mafua ai le oligonucleotides ma mononucleotides faʻatasi ma le 3'-OH ma le 5'-monophosphate pito, e le mafai ona faʻaleagaina le DNA tasi pe lua-stranded poʻo le RNA.

RNase

Galuega ma fa'aoga

E mafai e le Ribonuclease ona fa'atupuina le fa'aleagaina o le vaila'au ribonucleic (RNA) ma e mafai ona fa'apipi'iina fa'apitoa.O le suau'u fa'afoma'i e fa'aaogaina e togafitia ai manu'a ma tiga o so'oga.E tusa ai ma lipoti, e mafai e le ribonuclease ona suia le metabolism cell host, faʻalavelaveina le faʻapipiʻiina o siama, faʻalavelaveina le faʻateleina o le siama o le influenza in vitro, ma faʻalavelaveina le faʻavaeina o tui ma le herpes virus i totonu o moa moa.O le faʻaaogaina o le ribonuclease i aso taʻitasi tui intramuscular o le 180 mg, e aoga i le togafitiga o faʻamaʻi encephalitis, e mafai e le ribonuclease ona faʻaleagaina le faʻaleagaina o le ribonucleic acid (RNA), ma e mafai nei ona faʻapipiʻiina.O le suau'u fa'afoma'i e fa'aaogaina e togafitia ai manu'a ma tiga o so'oga.E tusa ai ma lipoti, e mafai e le ribonuclease ona suia le metabolism cell host, faʻalavelaveina le faʻapipiʻiina o siama, faʻalavelaveina le faʻateleina o le siama o le influenza in vitro, ma faʻalavelaveina le faʻavaeina o tui ma le herpes virus i totonu o moa moa.O le fa'aogaina o le ribonuclease i aso ta'itasi tui i totonu ole 180 mg e aoga mo le togafitia o fa'ama'i encephalitis.

RNase

Fa'auigaga fa'agata

Faʻamatalaga o le RNase inhibitor unit: Ole aofaʻi ole enzyme e manaʻomia e faʻalavelave ai le 50% o le gaioiga ole 5ng RNase A ole tasi le iunite.

Auala e galue ai RNase Inhibitors

Guanidine isothiocyanate:

Guanidine isothiocyanate ose fa'alapotopotoga fa'aola ma le fua mole mole C2H6N4S.E masani ona faʻaaogaina i vailaʻau faʻapitoa, vailaʻau faʻamaʻi, ma isi. Guanidine isothiocyanate o se vailaʻau lysing protein, ma e masani ona faʻaaogaina e avea ma vaega autu o le lysis solution i molecular diagnostic reagents.E mafai ona lyse aano, fa'aleagaina le fausaga o sela ma vavae ese le nucleic acid mai le nucleoprotein, ma e malosi le denaturation i le RNase.O le inhibitor RNase sili ona aoga i le taimi nei.

TRIzol o se tala fou atoa RNA extraction reagent e mafai ona aveese saʻo le RNA atoa mai sela poʻo masini.O loʻo i ai mea e pei o le phenol ma le guanidine isothiocyanate, lea e mafai ona faʻalavelave faʻafuaseʻi sela ma faʻalavelaveina nucleases e tuʻuina atu e sela.

(Ae ui i lea, o le guanidine isothiocyanate e matautia mo le soifua maloloina o tagata suʻesuʻe.)

RNasin:

Acid glycoprotein maua mai le ate iole po'o le blastoderm tagata.O le Rnasin o se inhibitor faʻatauvaʻa ole RNase, lea e mafai ona fusifusia i RNases eseese e faʻagata ai.

(Fa'amatalaga atili:https://www.foreivd.com/foreasy-rnase-inhibitor-product/)

Hmaualuga vevela:

O le maualuga o le vevela o se auala masani foi o le denaturation o porotini.

Diethylpyrocarbonate (DEPC):

O le DEPC o se inhibitor RNase malosi ae le atoatoa, lea e mafai ona faʻalavelaveina le gaioiga o le RNase e ala i le tuʻufaʻatasia ma le amino acid imidazole mama o le RNase vaega galue e denature proteins.

Vanadyl ribonucleoside lavelave:

O se faʻalavelave faʻapipiʻi e le vanadium oxide ions ma nucleosides, lea e fusifusia i le RNase i le tulaga o mea tau suiga, lea e toetoe lava a faʻalavelaveina le gaioiga a le RNase.

isi:

SDS, urea, diatomaceous earth, ma isi mea e iai foi se aafiaga faʻalavelave i le RNase.

Fa'amatalaga Fa'apitoa

Li Yujie Tekinisia Sili

Faʻatonu o le Matagaluega o Suʻesuʻega, Falemai Jiaozhou, Falemai Dongfang Faʻatasi i le Iunivesite o Tongji

Ina ia puipuia le biodegradation o le RNase mai fafo, matapulepule, totigilima ma pulou e tatau ona ofuina ma sui soo i le faagasologa o le aveeseina o le RNA.O ipu tioata uma e tatau ona tao i totonu o se ogaumu faʻamago i le 200°C mo le sili atu i le 2 itula.O mea e fa'aaogaina mo le taoina, e pei o palasitika, e mana'omia ona togafitia i le vai DEPC, ona fufulu lea i vai fa'amama.O reagents po'o masini e fa'aaogaina mo le fa'asaoina, fa'asaoina ma le fa'ailoaina o le RNA e tatau ona tu'uina atu i le RNA, ma e tatau ona fa'atūina se vaega tuto'atasi RNA fa'agaioiga.

fa'asinomaga:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.Fa'ama'i ma fa'ama'i i totonu o le nasopharynx a'o le'i alia'e mai fa'ama'i fa'ama'i tiga i lalo ole manava ile fanau a Ausetalia[J].Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Falema'i e Pulea Fa'ama'i Fa'ama'i Lala a le Fa'alapotopotoga Fa'afoma'i Puipuia Saina.Ta'iala mo le aoina ma le asiasia o fa'ata'ita'iga microbial falema'i [J].Tusitala Saina o Fa'ama'i Fa'ama'i, 2018(20):3192-3200.

[3] “Su'ega Fa'afoma'i Sefulu Afe Aiseā le Volume Su'esu'ega mole Biology"