Rnase A.
Ribonuclease A (RNaseA) ni polypeptide imwe imwe irimo imigozi 4 ya disulfide ifite uburemere bwa molekuline hafi 13.7 kDa.RNase Ais endoribonuclease itesha agaciro byumwihariko RNA imwe kumurongo wa C na U.By'umwihariko, clavage imenya isano ya fosifori yakozwe na 5'-ribose ya nucleotide hamwe nitsinda rya fosifate kuri 3'-ribose ya nucleotide ya pyrimidine yegeranye, kuburyo fosifeti 2,3'-Cyclic fosifate ihujwe na 3 ihuye 'nucleoside fosifate (urugero, pG-pG-pC-pA-pG yashizwemo na RNase A kugirango itange pG-pG-pCp na A-PG).RNase A nigikorwa cyane mugukata RNA imwe.Basabwe gukora cyane ni 1- 100 μ G / mL, bihujwe na sisitemu zitandukanye.Ubwinshi bwumunyu (0-100 mM NaCl) burashobora gukoreshwa mugukata RNA imwe, RNA ikubye kabiri, n'iminyururu ya RNA ikozwe na RNA-ADN ivangwa.
Nyamara, iyo umunyu mwinshi (≥0.3 M), RNase A ikuraho gusa RNA imwe.
RNase A ikoreshwa cyane mugukuraho RNA mugihe cyo gutegura ADN ya plasmide cyangwa ADN genomic.Niba DNase ikora cyangwa idakora mugihe cyo kwitegura irashobora guhindura byoroshye reaction.Uburyo gakondo bwo guteka mubwogero bwamazi burashobora gukoreshwa mugutangiza ibikorwa bya DNase.Ibicuruzwa ntabwo birimo DNase na protease, kandi ntibisaba kuvura ubushyuhe mbere yo kubikoresha.Mubyongeyeho, iki gicuruzwa kirashobora no gukoreshwa mubushakashatsi bwibinyabuzima bwa molekuline nko gusesengura kurinda RNase no gusesengura urutonde rwa RNA.
Imiterere yo kubika
Ibicuruzwa bigomba kubikwa kuri -25 ℃ ~ - 15 ℃ kumyaka 2.
Ibisobanuro
| Kugaragara | Ifu |
| Umubare | 100mg / 1g |
| Ubwoko bwibicuruzwa | RNase A. |
Amabwiriza
Ubu ni bumwe muburyo busanzwe bwo gutegura RNase Igisubizo cyo kubika.Irashobora kandi gutegurwanubundi buryo ukurikije uburyo gakondo muri laboratoire cyangwa ibitabo byerekana (nkagushonga mu buryo butaziguye muri 10 mM Tris-HCl, pH 7.5 cyangwa igisubizo cya Tris-NaCl)
1. Koresha 10 mM sodium acetate (pH 5.2) kugirango utegure 10 mg / mL ya RNase Igisubizo cyo kubika.
2. Gushyushya 100 ℃ kuminota 15.
3. Nkonje mubushyuhe bwicyumba, ongeramo 1/10 ingano ya 1 M Tris-HCl (pH 7.4), hindura pH kuri 7.4 (kuriurugero, ongeramo 500 mL ya 10 mg / mL Igisubizo cyo kubika RNase 1 M Tris-HCl, pH7.4).
4. Gupakira kuri -20 ℃ kubikwa bikonje, bishobora guhagarara neza kugeza kumyaka 2.
[Inyandiko]:
Iyo utetse igisubizo cya RNaseA mubihe bidafite aho bibogamiye, imvura ya RNase izashiraho;Guteka kuri pH yo hepfo, kandi niba hari imvura iguye, irashobora kugaragara, ishobora guterwa no kuba hariho proteine zanduye.Niba imyanda ibonetse nyuma yo gutekwa, umwanda urashobora gukurwaho na centrifugation yihuta (13000rpm), hanyuma ugashyirwa mububiko bwo gukonjesha.
Inyandiko
Nyamuneka kwambara PPE ikenewe, ikote rya laboratoire hamwe na gants, kugirango umenye ubuzima bwawe n'umutekano.
