Rnas A
Ribonuclease A (RNaseA) yog ib leeg-stranded polypeptide uas muaj 4 disulfide bonds nrog ib tug molecular hnyav li ntawm 13.7 kDa.RNase A yog ib qho endoribonuclease uas tshwj xeeb degrades ib leeg-stranded RNA ntawm C thiab U residues.Tshwj xeeb, qhov cleavage paub txog phosphodiester daim ntawv cog lus tsim los ntawm 5′-ribose ntawm ib nucleotide thiab phosphate pawg ntawm 3′-ribose ntawm cov pyrimidine nucleotide uas nyob ib sab, yog li ntawd 2, 3′-Cyclic phosphates yog hydrolyzed rau 3 sib xws. 'nucleoside phosphates (piv txwv li pG-pG-pC-pA-pG yog cleaved los ntawm RNase A los tsim pG-pG-pCp thiab A-PG).RNase A yog qhov tseem ceeb tshaj plaws hauv kev tshem tawm ib leeg-stranded RNA.Kev pom zoo ua haujlwm siab yog 1-100μG / mL, sib xws nrog ntau yam tshuaj tiv thaiv.Tsawg ntsev concentration (0-100 mM NaCl) tuaj yeem siv los txiav ib leeg-stranded RNA, ob-stranded RNA, thiab RNA chains tsim los ntawm RNA-DNA hybridization.Txawm li cas los xij, ntawm cov ntsev ntau (≥0.3 M), RNase A tsuas yog tshwj xeeb cleaves ib leeg-stranded RNA.
RNase A feem ntau yog siv los tshem tawm RNA thaum lub sijhawm npaj ntawm plasmid DNA lossis genomic DNA.Txawm tias DNase tsis ua haujlwm thaum lub sijhawm npaj tuaj yeem cuam tshuam cov tshuaj tiv thaiv.Cov txheej txheem ntawm boiling nyob rau hauv ib da dej yuav siv tau los inactivate DNase kev ua ub no.Cov khoom no tsis muaj DNase thiab protease, thiab tsis xav tau kev kho cua sov ua ntej siv.Tsis tas li ntawd, cov khoom no kuj tseem siv tau rau hauv kev sim molecular biology xws li RNase tiv thaiv kev tsom xam thiab RNA sequence tsom xam.
Kev cia khoom
Cov khoom tuaj yeem khaws cia ntawm -25 ~ 15 ℃, siv tau rau 2 xyoos.
Cov lus qhia
Qhov no yog ib txoj hauv kev los npaj RNase A cia tshuaj.Nws tseem tuaj yeem npaj los ntawmlwm txoj hauv kev raws li cov txheej txheem ib txwm siv hauv chav kuaj lossis cov ntaub ntawv siv (xws lincaj qha dissolving hauv 10 mM Tris-HCl, pH 7.5 los yog Tris-NaCl tov)
1. Siv 10 mM sodium acetate (pH 5.2) los npaj 10 mg/mL ntawm RNase A cia tshuaj
2. Cua sov ntawm 100 ℃ rau 15 min
3. Txias rau hauv chav sov, ntxiv 1/10 ntim ntawm 1 M Tris-HCl (pH 7.4), kho nws cov pH rau 7.4 (rauPiv txwv li, ntxiv 500 ml ntawm 10g / ml RNase cia tov 1M Tris-HCL, PH7.4)
4. Sub-packaged ntawm -20 ℃ rau khov cia, uas tuaj yeem ruaj khov mus txog 2 xyoos.
[Ceeb Toom]: Thaum boiling RNaseA tov nyob rau hauv nruab nrab tej yam kev mob, RNase nag yuav tsim;Boil nws ntawm qis pH, thiab yog tias muaj nag lossis daus, nws tuaj yeem pom, uas tuaj yeem tshwm sim los ntawm qhov muaj protein ntau impurities.Yog tias pom cov khib nyiab tom qab boiling, impurities tuaj yeem raug tshem tawm los ntawm kev kub ceev centrifugation (13000rpm), thiab ces sub-packed rau khov cia.
Cov ntaub ntawv khoom
| Synonyms | Ribonuclease kuv;Pancreatic ribonuclease;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease kuv |
| CAS Nr. | 9001-99-4 |
| Qhov tshwm sim | Dawb lyophilized hmoov |
| Molecular hnyav | ~ 13.7kDa (amino acid sequence) |
| Ph nqi | 7.6 (ua haujlwm ntau 6-10) |
| Haum Kub | 60 ℃ (Kev ua haujlwm ntau 15-70 ℃) |
| Activating agent | Na2+.K+ |
| Inhibitor | Rnase Inhibitor |
| Txoj kev Inactivation | Tsis tuaj yeem ua haujlwm los ntawm cua sov, nws raug nquahu kom siv kab centrifuge |
| Keeb kwm | Bovine |
| Solubility | Soluble hauv dej (10mg / ml) |
| Poob rau qhuav | ≤ 5.0% |
| Kev ua haujlwm enzyme | ≥60 Knitz units / mg |
| Isoelectric point | 9.6 ib |
Sau ntawv
Rau koj txoj kev nyab xeeb thiab kev noj qab haus huv, thov hnav lub tsho (lab) thiab cov hnab looj tes pov tseg rau kev ua haujlwm.
