Abanditsi: Wang Xiaoyan, Zhao Eryu

Igice: Ibitaro bya Jiaozhou, Ibitaro bya Dongfang Bishamikiye kuri kaminuza ya Tongji

Kugeza ubu, ubwoko bw'icyitegererezo cy'ubuhumekero butera aside nucleic aside ni umuhogo.Hariho ubwoko bwinshi bwibisanzwe bikoreshwa muburyo bwo kubika ibisubizo, byose bigomba gukonjeshwa cyangwa gukonjeshwa kugirango bitwarwe nububiko;Biragoye kugenzura inzira yose yubushyuhe buke mugihe cyo gukusanya no gutwara, kandi biragoye kwemeza ubuziranenge mbere yo gupima icyitegererezo^[1-2].

RNase (RNase) ni endonuc Please hydrolyzes RNA, cyane cyane igabanya isano ya fosifori hagati ya nucleotide.Molekile ya RNase irahagaze neza, hariho imiyoboro ya disulfide mumiterere, kandi ibikorwa byayo ntibisaba ko habaho cations zingana, bityo rero RNase ntabwo yangwa byoroshye, kandi biroroshye guhinduka nubwo nyuma yubushyuhe bwinshi cyangwa gukoresha denaturants.RNase igabanijwemo endogenous na exogenous.Endogenous RNase irashobora kurekurwa mugihe selile zimaze guturika.Kubwibyo, gukuraho uruhare rwa endogenous RNase nintambwe ikomeye cyane mugikorwa cyo gukuramo RNA.Exogenous RNase ikwirakwizwa cyane.RNase ibaho mu kirere, uruhu rwabantu, umusatsi nu macandwe, nizo mpamvu zingenzi zo kwangirika byoroshye kwa RNA^[3].

Ikibazo

CANS-CL02-A009 “Gushyira mu bikorwa Amabwiriza ya Laboratoire y’ubuvuzi n’ubushobozi bwo kwemeza ubushobozi mu rwego rwo gusuzuma indwara ya molekuline” mu bisabwa bya tekiniki avuga ko hagomba gushyirwaho ibipimo ngenderwaho by’amazi bikwiye hakurikijwe ibisabwa;Ikoreshwa ryumuyaga mwinshi:

RNase / Ibyiciro

(1) RNase A.

Ribonuclease A (RNase A), ikomoka kuri bovine pancreas, ni endoribonuclease ishobora kwibasira cyane 3 ′ iherezo ryibisigisigi bya pyrimidine kuri RNA, ikata cytosine cyangwa uracil ikorwa na nucleotide yegeranye.Inkunga ya Phosphodiester, igicuruzwa cya nyuma cyibisubizo ni 3 ′ pyrimidine nucleotide na oligonucleotide hamwe na 3 ′ pyrimidine nucleotide irangiye.

(2) RNase T1

Ribonuclease T1 (RNase T1) yakomotse kuri Aspergillus orjzae, ikora cyane cyane kuri fosifate ya 3′-ya fosifate ya guanine, kandi ikibanza cya clavage kiri hagati ya 3 ′ fosifate ya guanine na 5 ′ hydroxyl ya nucleotide yegeranye.Igicuruzwa cyanyuma cyibisubizo ni 3′guanylic aside hamwe na oligonucleotide ibice hamwe na 3′guanylic aside irangiye.

(3) RNase H.

Ribonuclease H (RNase H) yavumbuwe bwa mbere mu nyana ya thymus yinyana, kandi gene yayo yashizwemo yashyizwe muri coli ya Escherichia.Irashobora gutesha agaciro ADN cyane: imirongo ya RNA muri duplex ya RNA, bikavamo oligonucleotide na mononucleotide hamwe na 3′-OH na 5′-monofosifate, ntishobora gutesha agaciro ADN imwe cyangwa ebyiri.

RNase

Imikorere no gukoresha

Ribonuclease irashobora guhagarika iyangirika rya aside ribonucleic (RNA) kandi irashobora guhuzwa muburyo bwa artile.Amavuta yimiti akoreshwa cyane mukuvura ihahamuka nububabare.Nk’uko amakuru abitangaza, ribonuclease irashobora guhindura metabolisme yakira, ikabuza kwanduza virusi, ikabuza ikwirakwizwa rya virusi yibicurane muri vitro, kandi ikabuza kwandura virusi na herpes mu nsoro z’inkoko.Gukoresha ivuriro rya ribonuclease buri munsi inshinge za mg 180, zifasha kuvura icyorezo cya encephalitis, ribonuclease irashobora gutuma iyangirika rya acide ribonucleic (RNA), kandi ubu irashobora guhuzwa.Amavuta yimiti akoreshwa cyane mukuvura ihahamuka nububabare.Nk’uko amakuru abitangaza, ribonuclease irashobora guhindura metabolisme yakira, ikabuza kwanduza virusi, ikabuza ikwirakwizwa rya virusi yibicurane muri vitro, kandi ikabuza kwandura virusi na herpes mu nsoro z’inkoko.Gukoresha ivuriro rya ribonuclease buri munsi inshinge zo mu nda ya mg 180 ni ingirakamaro mu kuvura icyorezo cya encephalite.

RNase

Inhibitor Ibisobanuro

Igisobanuro cya RNase inhibitor unit: Ingano ya enzyme isabwa kugirango ibuze 50% yibikorwa bya 5ng RNase A nigice kimwe.

Uburyo RNase Inhibitor ikora

Guanidine isothiocyanate:

Guanidine isothiocyanate ni ifumbire mvaruganda hamwe na molekile ya C2H6N4S.Ahanini ikoreshwa mubuvuzi bwibinyabuzima, reagent ya chimique, nibindi. Guanidine isothiocyanate ni proteine ​​lysing agent, kandi akenshi ikoreshwa nkigice cyingenzi cyibisubizo bya lysis muri reagent yo kwisuzumisha.Irashobora gutera ingirabuzimafatizo, gusenya imiterere ya selile no gutandukanya aside nucleic na nucleoproteine, kandi ikagira ihinduka rikomeye kuri RNase.Nibikorwa byiza bya RNase inhibitor muri iki gihe.

TRIzol nigitabo gishya cyo gukuramo RNA gishobora gukuramo RNA yose muri selile cyangwa tissue.Irimo ibintu nka fenol na guanidine isothiocyanate, bishobora guhungabanya byihuse ingirabuzimafatizo no kubuza nuclease kurekurwa na selile.

(Nyamara, guanidine isothiocyanate ni mbi kubuzima bwabashakashatsi ba laboratoire.)

RNasin:

Acide glycoproteine ​​yakuwe mu mwijima wimbeba cyangwa blastoderm yumuntu.Rnasin ni inzitizi idahwitse ya RNase, ishobora guhuza RNase zitandukanye kugirango idakora.

(Ibisobanuro birambuye: https: //www.foreivd.com/impamvu-rnase-inhibitor-product/)

High ubushyuhe:

Ubushyuhe bwo hejuru nabwo ni uburyo busanzwe bwo gutandukanya poroteyine.

Diethylpyrocarbonate (DEPC):

DEPC ni inhibitori ya RNase ikomeye ariko ituzuye, irashobora guhagarika ibikorwa bya RNase muguhuza impeta ya amino acide imidazole impeta yitsinda rya RNase kugirango yamagane poroteyine.

Vanadyl ribonucleoside complex:

Urusobekerane rwakozwe na vanadium oxyde ion na nucleoside, ihuza na RNase muburyo bwinzibacyuho, ishobora guhagarika rwose ibikorwa bya RNase

ibindi:

SDS, urea, isi ya diatomaceous, nibindi nabyo bigira ingaruka runaka kuri RNase.

Isuzuma ry'impuguke

Li Yujie Umutekinisiye mukuru

Umuyobozi w'ishami rya Laboratoire, Ibitaro bya Jiaozhou, Ibitaro bya Dongfang Bishamikiye kuri kaminuza ya Tongji

Kugirango wirinde kwangiza ibinyabuzima bya RNase idasanzwe, masike, gants n'ingofero bigomba kwambarwa no gusimburwa kenshi mugihe cyo gukuramo RNA.Ibirahuri byose bigomba gutekwa mu ziko ryumye kuri 200 ° C mumasaha arenze 2.Ibikoresho bikoreshwa muguteka, nka plastiki, bigomba kuvurwa namazi ya DEPC, hanyuma bigakaraba namazi yatoboye.Reagent cyangwa ibikoresho bikoreshwa mugukuramo, kubungabunga no kumenyekanisha RNA bigomba kuba byeguriwe RNA, kandi hagomba gushyirwaho agace kigenga ka RNA.

Reba:

[1] Smith-Vaughan HC, Binks MJ, Beissbarth J, n'abandi.Indwara ya bagiteri na virusi muri nasofarynx ako kanya mbere yo gutangira kwandura indwara zikomeye z’ubuhumekero ku bana b’abasangwabutaka bo muri Ositaraliya [J].Eur J Clin Microbiol Yanduye Disiki, 2018,37 (9): 1785-1794.

Ishami rishinzwe kurwanya indwara z’ibitaro ry’ishyirahamwe ry’ubuvuzi bukumira abashinwa.Amabwiriza yo gukusanya no kugenzura imiterere ya mikorobe ivura [J].Ikinyamakuru cyo mu Bushinwa cyanduye ibitaro, 2018 (20): 3192-3200.

[3] “Ikizamini cya Clinical Ibihumbi icumi Kuki Ikizamini cya Molecular Biologiya”