Bangoli: Wang Xiaoyan, Zhao Eryu

Yuniti: Sepetlele sa Jiaozhou, Sepetlele sa Dongfang se amanang le Univesithi ea Tongji

Hajoale, mofuta oa mantlha oa sampole bakeng sa ho lemoha pathogen nucleic acid ke 'metso.Ho na le mefuta e mengata ea litharollo tse sebelisoang ka tloaelo tsa ho boloka mohlala, tseo kaofela li hlokang ho beoa ka sehatsetsing kapa ho hoamisoa bakeng sa lipalangoang le polokelo;Ho thata ho laola ts'ebetso eohle ea mocheso o tlase nakong ea pokello le ts'ebetso ea lipalangoang, 'me ho thata ho netefatsa taolo ea boleng pele ho tlhahlobo ea sampole.^[1-2].

RNase (RNase) ke endonuclease e etsang hydrolyzes RNA, haholo-holo e khaola maqhama a phosphodiester lipakeng tsa li-nucleotide.Molek'hule ea RNase e tsitsitse haholo, ho na le litlamo tsa disulfide mohahong, 'me mosebetsi oa eona ha o hloke ho ba teng ha li-cations tsa divalent, kahoo RNase ha e fokotsehe habonolo,' me ho bonolo ho fetola bocha esita le ka mor'a mocheso o phahameng kapa tšebeliso ea li-denaturants.RNase e arotsoe ka endogenous le exogenous.Endogenous RNases e ka lokolloa ka nako e le 'ngoe ha lisele li phatlohile.Ka hona, ho felisa karolo ea li-RNase tse sa feleng ke mohato oa bohlokoa haholo ts'ebetsong ea ho ntša RNA.Li-RNase tsa Exogenous li ajoa haholo.Li-Rnase li teng moeeng, letlalong la motho, moriri le mathe, e leng mabaka a bohlokoa a ho senyeha habonolo ha RNA.^[3].

Potso ea data

CANS-CL02-A009 "Kopo ea Litaelo tsa Kamohelo ea Boleng ba Laboratori ea Bongaka le Bokhoni Lefapheng la Tlhahlobo ea Limolek'hule" ka litlhoko tsa tekheniki e fana ka maikutlo a hore litekanyetso tse nepahetseng tsa boleng ba metsi li lokela ho etsoa ho latela kopo;Lijana tse lahloang tse sa keneng moea:

RNase/ Ketso

(1) RNase A

Ribonuclease A (RNase A), e tsoang ho li-pancreas tsa bovine, ke endoribonuclease e ka hlaselang ka ho khetheha karolo ea 3 ea masala a pyrimidine ho RNA, ho khaola cytosine kapa uracil e entsoeng ke li-nucleotide tse haufi.Phosphodiester bond, sehlahisoa sa ho qetela sa karabelo ke 3 'pyrimidine nucleotide le oligonucleotide e nang le 3' pyrimidine nucleotide qetellong.

(2) RNase T1

Ribonuclease T1 (RNase T1) e nkiloe ho Aspergillus orjzae, ka ho khetheha e sebetsa ho 3′-terminal phosphate ea guanine, 'me sebaka sa cleavage se pakeng tsa 3′ phosphate of guanine le 5′ hydroxyl ea nucleotide e haufi.Sehlahisoa sa ho qetela sa karabelo ke 3′guanylic acid le likhechana tsa oligonucleotide tse nang le 3′guanylic acid qetellong.

(3) RNase H

Ribonuclease H (RNase H) e ile ea fumanoa ka lekhetlo la pele ho tsoa ho namane ea thymus, 'me mofuta oa eona oa khouto o entsoe ka Escherichia coli.E ka senya DNA ka ho khetheha: likhoele tsa RNA ho li-duplexes tse nyalisitsoeng tsa RNA, tse hlahisang oligonucleotides le mononucleotide tse nang le 3'-OH le 5'-monophosphate lipheletsong, e ke ke ea senya DNA kapa RNA e le 'ngoe kapa habeli.

RNase

Mosebetsi le tšebeliso

Ribonuclease e ka baka ho senyeha ha ribonucleic acid (RNA) mme e ka etsoa ka maiketsetso.Setlolo sa meriana se sebelisoa ka holimo ho phekola leqeba le bohloko ba manonyeletso.Ho ea ka litlaleho, ribonuclease e ka fetola metabolism ea lisele tse amohelang, ea thibela kokoana-hloko synthesis, ea thibela ho ata ha kokoana-hloko ea ntaramane ka har'a vitro, 'me ea thibela ho thehoa ha vaccinia le kokoana-hloko ea herpes ka mahe a mahe a likhoho.Tšebeliso ea bongaka ea letsatsi le letsatsi ea ribonuclease intramuscular ente ea 180 mg, e molemo bakeng sa phekolo ea lefu la seoa la encephalitis, ribonuclease e ka baka ho senyeha ha ribonucleic acid (RNA), 'me joale e ka etsoa ka maiketsetso.Setlolo sa meriana se sebelisoa ka holimo ho phekola leqeba le bohloko ba manonyeletso.Ho ea ka litlaleho, ribonuclease e ka fetola metabolism ea lisele tse amohelang, ea thibela kokoana-hloko synthesis, ea thibela ho ata ha kokoana-hloko ea ntaramane ka har'a vitro, 'me ea thibela ho thehoa ha vaccinia le kokoana-hloko ea herpes ka mahe a mahe a likhoho.Tšebeliso ea bongaka ea ribonuclease letsatsi le leng le le leng ea intramuscular ea 180 mg e molemo bakeng sa phekolo ea lefu la seoa la encephalitis.

RNase

Inhibitor Tlhaloso

Tlhaloso ea RNase inhibitor unit: Palo ea enzyme e hlokahalang ho thibela 50% ea mosebetsi oa 5ng RNase A ke yuniti e le 'ngoe.

Kamoo RNase Inhibitors e sebetsang kateng

Guanidine isothiocyanate:

Guanidine isothiocyanate ke motsoako oa manyolo o nang le foromo ea molek'hule C2H6N4S.Haholo-holo e sebelisoa meriana ea likokoana-hloko, li-reagents tsa lik'hemik'hale, joalo-joalo Guanidine isothiocyanate ke protheine lysing agent, 'me e atisa ho sebelisoa e le karolo e ka sehloohong ea tharollo ea lysis ho li-reagents tsa ho hlahloba limolek'hule.E ka senya lisele, ea senya sebopeho sa lisele le ho arola nucleic acid ho nucleoprotein, 'me e na le denaturation e matla ho RNase.Ke RNase inhibitor e sebetsang ka ho fetisisa hajoale.

TRIzol ke "Reagent" ea "RNA" e ncha e ka ntšang RNA e felletseng liseleng kapa liseleng.E na le lintho tse kang phenol le guanidine isothiocyanate, tse ka senyang lisele ka potlako le ho thibela li-nuclease tse lokolloa ke lisele.

(Leha ho le joalo, guanidine isothiocyanate e kotsi bakeng sa bophelo bo botle ba bafuputsi ba lab.)

RNasin:

Acid glycoprotein e ntšitsoeng sebeteng sa rat kapa blastoderm ea motho.Rnasin ke inhibitor e seng tlholisano ea RNase, e ka tlamang li-RNase tse fapaneng ho li etsa hore li sebetse.

(Lintlha tse ling: https://www.foreivd.com/foreasy-rnase-inhibitor-product/)

Hmocheso o phahameng:

Mocheso o phahameng hape ke mokhoa o tloaelehileng oa denaturation ea protheine.

Diethylpyrocarbonate (DEPC):

DEPC ke RNase inhibitor e matla empa e sa fella, e ka thibelang ts'ebetso ea RNase ka ho kopanya le reng ea imidazole ea amino acid ea sehlopha se sebetsang sa RNase ho hlahisa liprotheine.

Vanadyl ribonucleoside complex:

Motsoako o entsoeng ke vanadium oxide ion le nucleosides, o tlamahanyang le RNase ka mokhoa oa lintho tse fetohang, tse ka batlang ho thibela tšebetso ea RNase ka botlalo.

tse ling:

SDS, urea, diatomaceous earth, joalo-joalo le tsona li na le phello e itseng ea thibelo ho RNase.

Litlhahlobo tsa Setsebi

Setsebi se ka Sehloohong sa Li Yujie

Motsamaisi oa Lefapha la Laboratori, Sepetlele sa Jiaozhou, Sepetlele sa Dongfang se amanang le Univesithi ea Tongji

E le ho thibela ho senyeha ha likokoana-hloko tsa RNase e sa tloaelehang, limaske, liatlana le likatiba li lokela ho apesoa le ho nkeloa sebaka khafetsa nakong ea ts'ebetso ea ho ntša RNA.Lisebelisoa tsohle tsa khalase li tlameha ho behoa ka ontong e omisang ka 200 ° C bakeng sa lihora tse fetang 2.Lisebelisoa tse sebelisetsoang ho baka, tse kang polasetiki, li hloka ho phekoloa ka metsi a DEPC, ebe li hlatsuoa ka metsi a silafetseng.Lisebelisoa kapa lisebelisoa tse sebelisoang bakeng sa ho ntša, ho boloka le ho tsebahatsa RNA li lokela ho neheloa ho RNA, 'me sebaka se ikemetseng sa ts'ebetso ea RNA se lokela ho thehoa.

litšupiso:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, et al.Likokoana-hloko le livaerase ka har'a nasopharynx hang pele ho qala mafu a matla a tlase a ho hema ho bana ba Maindia a Australia[J].Eur J Clin Microbiol Infect Dis,2018,37 (9): 1785-1794.

[2] Lekala la Taolo ea Tšoaetso ea Lipetlele la Mokhatlo oa Chaena oa Thibelo ea Meriana.Litaelo tsa ho bokella le ho hlahloba mehlala ea likokoana-hloko tsa kliniki [J].Chinese Journal of Hospital Infection, 2018(20):3192-3200.

[3] “Teko ea Lingaka Tse Likete Tse Leshome Ke Hobane’ng ha Bolumo ea Teko ea Limolek’hule”