Rnase A
Ribonuclease A(RNaseA) ke polypeptide e nang le khoele e le 'ngoe e nang le li-bond tse 4 tsa disulfide tse nang le boima ba molek'hule bo ka bang 13.7 kDa.RNase Ais ke endoribonuclease e nyenyefatsang ka ho khetheha RNA ea makhalo a le mong ho masala a C le U.Haholo-holo, 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 le 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- 100 μ G/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, ha letsoai le le lengata (≥0.3 M), RNAse A e petsola feela RNA e nang le thipa e le 'ngoe.
RNase A e sebelisoa haholo ho tlosa RNA nakong ea ho lokisoa ha plasmid DNA kapa genomic DNA.Hore na DNase e sebetsa kapa che nakong ea ts'ebetso 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 lokela ho bolokoa ho -25 ℃ ~ - 15 ℃ ka lilemo tse 2.
Litlhaloso
| Ponahalo | Phofo |
| Bongata | 100mg / 1g |
| Mofuta oa sehlahisoa | RNase A |
Litaelo
Ena ke e 'ngoe ea mekhoa e tloaelehileng ea ho lokisa tharollo ea polokelo ea RNase A.E ka boela ea lokisoaka mekhoa e meng ho latela mekhoa ea khale ea laboratori 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 tharollo ea polokelo ea RNase A.
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 10 mg/mL RNase polokelo tharollo 1 M 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 fumanwa ka mora ho bela, ditshila di ka tloswa ka lebelo le phahameng la centrifugation (13000rpm), ebe di pakwa ka tlase bakeng sa polokelo ya serame.
Lintlha
Ka kopo, roala PPE e hlokahalang, jase e joalo ea lab le liatlana, ho netefatsa bophelo bo botle le polokeho ea hau.
