Rnase A.
Ribonuclease A (RNaseA) ni polypeptide imwe imwe irimo imigozi 4 ya disulfide ifite uburemere bwa molekile hafi 13.7 kDa.RNase A ni endoribonuclease itesha agaciro cyane RNA imwe imwe kuri 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′-Cyclica ziba hydrolyz kuri 3 ihuye na 3 Os nucleoside fosifate (urugero: pG-pG-pC-pA-pG ikozwe na RNase A kugirango itange pG-pG-pCp na A-PG).RNase A nigikorwa cyane mugukata RNA imwe.Basabwe gukora cyane ni 1-100μG / 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 ya genomique.Nubwo DNase ikora cyangwa idakora mugihe cyimyiteguro irashobora kugira ingaruka kubitekerezo.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 gishobora no gukoreshwa mubushakashatsi bwibinyabuzima bwa molekuline nko gusesengura kurinda RNase no gusesengura urutonde rwa RNA.
Ububiko
Ibicuruzwa birashobora kubikwa kuri -25 ～ -15 ℃, byemewe kumyaka 2.
Amabwiriza
Ubu ni bumwe muburyo busanzwe bwo gutegura RNase Igisubizo cyo kubika.Irashobora kandi gutegurwa naubundi 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 ml 500 ya 10g / ml Igisubizo cyo kubika RNase 1M Tris-HCL, PH7.4)
4. Gupakira kuri -20 ℃ kubikwa bikonje, bishobora guhagarara neza kugeza kumyaka 2.
[Icyitonderwa]: 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.
Amakuru y'ibicuruzwa
| Synonyme | Ribonuc Please I;Pancreatic ribonuclease;Ribonuc Please 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I. |
| URUBANZA No. | 9001-99-4 |
| Kugaragara | Ifu yera ya lyofilize |
| Uburemere bwa molekile | ~ 13.7kDa (urukurikirane rwa aside amine) |
| Agaciro Ph | 7.6 (urutonde rwibikorwa 6-10) |
| Ubushyuhe bukwiye | 60 ℃ (Ibikorwa biri hagati ya 15-70 ℃) |
| Umukozi ukora | Na2+.K.+ |
| Inhibitor | Rnase Inhibitor |
| Uburyo bwo kudakora | Ntushobora kudakorwa no gushyushya, birasabwa gukoresha centrifuge |
| Inkomoko | Bovine |
| Gukemura | Gushonga mumazi (10mg / ml) |
| Gutakaza byumye | ≤5.0% |
| Igikorwa cya Enzyme | Units60 Kunitz ibice / mg |
| Ingingo ya Isoelectric | 9.6 |
Inyandiko
Kubwumutekano wawe nubuzima bwawe, nyamuneka wambare amakoti ya laboratoire hamwe na gants imwe ikoreshwa.
