Rnase A
Ribonuclease A (RNaseA) ke polypeptide e nang le khoele e le 'ngoe e nang le li-bond tsa 4 tsa disulfide tse nang le boima ba molek'hule bo ka bang 13.7 kDa.RNase A ke endoribonuclease e nyenyefatsang ka ho khetheha RNA ea makhalo a le mong ho masala a C le U.Ka ho khetheha, cleavage e lemoha tlamo ea phosphodiester e entsoeng ke 5'-ribose ea nucleotide le sehlopha sa phosphate holim'a 3'-ribose ea pyrimidine nucleotide e haufi, e le hore 2, 3'-Cyclic phosphates e be hydrolyzed ho 3 e lumellanang. "nucleoside phosphates (mohlala, pG-pG-pC-pA-pG e petsotsoe ke RNase A ho hlahisa pG-pG-pCp le A-PG).RNase A ke eona e sebetsang ka ho fetisisa ho petsola RNA e nang le thipa e le 'ngoe.Khokahano ea ts'ebetso e khothalelitsoeng ke 1-100μG/mL, e tsamaellana le litsamaiso tse fapaneng tsa karabelo.Letsoai le tlaase (0-100 mM NaCl) le ka sebelisoa ho khaola RNA e nang le likhoele tse le 'ngoe, RNA e nang le likarolo tse peli, le liketane tsa RNA tse entsoeng ke RNA-DNA hybridization.Leha ho le joalo, ka har'a letsoai le lengata (≥0.3 M), RNase A e petsola feela RNA ea thipa e le 'ngoe.
RNase A e atisa ho sebelisoa ho tlosa RNA nakong ea ho lokisoa ha plasmid DNA kapa genomic DNA. Ho sa tsotellehe hore na DNase e sebetsa kapa che nakong ea ho itokisa e ka ama karabelo habonolo.Mokhoa o tloaelehileng oa ho belisa ka bateng ea metsi o ka sebelisoa ho tima ts'ebetso ea DNase.Sehlahisoa sena ha se na DNase le protease, 'me ha se hloke phekolo ea mocheso pele se sebelisoa.Ntle le moo, sehlahisoa sena se ka sebelisoa litekong tsa baeloji ea limolek'hule joalo ka tlhahlobo ea ts'ireletso ea RNase le tlhahlobo ea tatelano ea RNA.
Maemo a polokelo
Sehlahisoa se ka bolokoa ho -25～-15℃, se sebetsa lilemo tse peli.
Litaelo
Ena ke e 'ngoe ea mekhoa e tloaelehileng ea ho lokisa tharollo ea polokelo ea RNase A.E ka boela ea lokisoa kemekhoa e meng ho latela mekhoa ea khale ka laboratoring kapa lingoliloeng tsa litšupiso (joalo kaho qhibiliha ka kotloloho ho 10 mM Tris-HCl, pH 7.5 kapa tharollo ea Tris-NaCl)
1. Sebelisa 10 mM sodium acetate (pH 5.2) ho lokisetsa 10 mg/mL ea RNase A polokelo tharollo.
2. Ho futhumatsa ka 100 ℃ bakeng sa 15 min
3. Pholile ho mocheso oa kamore, eketsa 1/10 molumo oa 1 M Tris-HCl (pH 7.4), fetola pH ea eona ho 7.4 (bakeng samohlala, eketsa 500 ml ea 10g/ml RNase polokelo tharollo 1M Tris-HCL, PH7.4)
4. E kentsoe ka har'a -20℃ bakeng sa polokelo e hatselitsoeng, e ka tsitsang ho fihlela lilemo tse 2.
[Lintlha]: Ha u belisa tharollo ea RNaseA tlas'a maemo a sa nke lehlakore, pula ea RNase e tla theha;E belise ka pH e tlase, 'me haeba ho na le pula, e ka bonoa, e ka bakoang ke boteng ba litšila tsa protheine.Haeba sediment e ka fumanoa ka mor'a ho bela, litšila li ka tlosoa ka lebelo le phahameng la centrifugation (13000rpm), ebe li kenngoa ka tlaase bakeng sa polokelo ea leqhoa.
Lintlha tsa sehlahisoa
| Litlhaloso tse tšoanang | Ribonuclease I;pancreatic ribonuclease;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I |
| Nomoro ea CAS. | 9001-99-4 |
| Ponahalo | White lyophilized phofo |
| Boima ba molek'hule | ~ 13.7kDa (tatellano ea amino acid) |
| Ph Boleng | 7.6 (ketso ea 6-10) |
| Mocheso o Loketseng | 60 ℃ (Lebaka la ts'ebetso 15-70 ℃) |
| Moemedi wa tshebetso | Na2+.K+ |
| Inhibitor | Rnase Inhibitor |
| Mokhoa oa ho kenya tšebetsong | Ha e khone ho tima ka ho futhumatsa, ho kgothaletswa ho sebedisa kholomo ya centrifuge |
| Tšimoloho | Bovine |
| Solubility | E qhibiliha ka metsing (10mg/ml) |
| Tahlehelo ha o omile | ≤5.0% |
| Ts'ebetso ea enzyme | ≥60 Kunitz diyuniti/mg |
| Ntlha ea Isoelectric | 9.6 |
Lintlha
Bakeng sa polokeho le bophelo bo botle ba hau, ka kopo roala lijase tsa lab le liatlana tse lahloang bakeng sa ts'ebetso.
