Rnase A
Ribonuclease A (RNaseA) bụ polypeptide nwere otu eriri nwere njikọ disulfide 4 nwere ịdị arọ molekụla nke ihe dịka 13.7 kDa.RNase A bụ endoribonuclease nke na-eweda RNA nwere otu eriri na ihe fọdụrụ C na U.Kpọmkwem, nkewa ahụ na-amata njikọ nke phosphodiester nke 5'-ribose nke nucleotide na otu phosphate na 3'-ribose nke nucleotide pyrimidine dị n'akụkụ, nke mere na 2,3'-Cyclic phosphates na-hydrolyzed na 3 kwekọrọ. 'nucleoside phosphates (dịka, pG-pG-pC-pA-pG bụ RNase A gbawara iji mepụta pG-pG-pCp na A-PG).RNase A bụ nke kacha arụsi ọrụ ike na ikpochapụ RNA nwere otu eriri.Ntinye uche na-arụ ọrụ akwadoro bụ 1-100 μG / ml, dakọtara na sistemụ mmeghachi omume dị iche iche.Enwere ike iji ntinye nnu dị ala (0-100 mM NaCl) belata RNA nwere eriri, RNA nwere eriri abụọ, na ụdọ RNA nke RNA-DNA kpụrụ.
Agbanyeghị, na ntinye nnu dị elu (≥0.3 M), RNase A na-ekewa RNA nwere otu eriri.
A na-ejikarị RNase A wepụ RNA n'oge nkwadebe nke DNA plasmid ma ọ bụ DNA genomic.Ma DNAse adịghị arụ ọrụ n'oge usoro nkwadebe nwere ike imetụta mmeghachi omume ahụ n'ụzọ dị mfe.Enwere ike iji usoro ọdịnala nke esi esi n'ime mmiri ịsa ahụ mee ka ọrụ DNAse rụọ ọrụ.Ngwaahịa a enweghị DNAase na protease, ọ dịghịkwa achọ ọgwụgwọ okpomọkụ tupu eji ya.Na mgbakwunye, enwere ike iji ngwaahịa a na nnwale ihe omimi ihe omimi dị ka nyocha nchekwa RNase na nyocha usoro RNA.
Ọnọdụ nchekwa
Ekwesịrị ịchekwa ngwaahịa ahụ na -25 ℃ ~ - 15 ℃ maka afọ 2.
Nkọwapụta
| Ọdịdị | Ntụ ntụ |
| Ọnụọgụ | 100mg / 1g |
| Ụdị ngwaahịa | RNase A |
Ntuziaka
Nke a bụ otu n'ime ụzọ a na-ejikarị akwadebe ihe nchekwa nchekwa RNase A.Enwere ike ịkwado yasite na ụzọ ndị ọzọ dịka usoro ọdịnala si dị na ụlọ nyocha ma ọ bụ akwụkwọ ntụaka (dịkana-agbaze ozugbo na 10 mM Tris-HCl, pH 7.5 ma ọ bụ ngwọta Tris-NaCl)
1. Jiri 10 mM sodium acetate (pH 5.2) kwadebe 10 mg / mL nke RNase ngwọta nchekwa.
2. Na-ekpo ọkụ na 100 ℃ maka 15 min.
3. Cool ka ụlọ okpomọkụ, tinye 1/10 olu nke 1 M Tris-HCl (pH 7.4), gbanwee pH ya na 7.4 (makaọmụmaatụ, tinye 500 ml nke 10 mg/mL RNase ngwọta nchekwa 1 M Tris-HCl, pH7.4).
4. Sub-kwakọtara na -20 ℃ maka oyi kpọnwụrụ nchekwa, nke nwere ike kwụsie ike ruo 2 afọ.
[Ihe edeturu]:
Mgbe ị na-esi nri RNaseA n'okpuru ọnọdụ nnọpụiche, mmiri ozuzo RNase ga-etolite;Sichaa ya na pH dị ala, ma ọ bụrụ na enwere mmiri ozuzo, enwere ike ịhụ ya, nke nwere ike ime site na ọnụnọ protein adịghị ọcha.Ọ bụrụ na ahụrụ sedimenti ka esichara, enwere ike wepu ihe adịghị ọcha site na centrifugation dị elu (13000rpm), wee kwakọba ya maka nchekwa oyi.
Ihe ndetu
Biko yiri PPE dị mkpa, uwe ụlọ nyocha na uwe aka, iji hụ na ahụike na nchekwa gị.
