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, mgbawa 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 1-100μG / 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 na-ewepụ RNA n'oge nkwadebe nke DNA plasmid ma ọ bụ DNA genomic. Ma DNA na-arụ ọrụ n'oge usoro nkwadebe nwere ike imetụta mmeghachi omume 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 nyocha ihe ọmụmụ ihe omimi dị ka nyocha nchekwa RNase na nyocha usoro RNA.
Ọnọdụ Nchekwa
Enwere ike ịchekwa ngwaahịa ahụ na -25 ~ -15 ℃, dị irè maka afọ 2.
Ntuziaka
Nke a bụ otu n'ime ụzọ a na-ejikarị akwadebe ihe nchekwa nchekwa RNase A.Enwere ike ịkwadebe ya site naụzọ ndị ọzọ dị ka 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 10g/ml RNase ngwọta nchekwa 1M Tris-HCL, PH7.4)
4. Sub-kwakọrọ 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, oke mmiri 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.
Ozi ngwaahịa
| Ụdị okwu | Ribonuclease I;Pancreatic ribonuclease;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I |
| CAS Mba. | 9001-99-4 |
| Ọdịdị | ntụ ntụ lyophilized ọcha |
| Ibu molekụla | ~ 13.7kDa (usoro amino acid) |
| Ph Uru | 7.6 (Oke ọrụ 6-10) |
| Okpomọkụ dabara adaba | 60℃ (Oghere ọrụ 15-70℃) |
| Onye na-arụ ọrụ | Na2+.K+ |
| Ihe mgbochi | Ihe mgbochi Rnase |
| Usoro adịghị arụ ọrụ | Enweghị ike ịmegharị ya site na kpo oku, a na-atụ aro ka iji kọlụm centrifuge |
| Mmalite | Anụ anụ |
| Solubility | Soluble na mmiri (10mg / ml) |
| Ọnwụ na akọrọ | ≤5.0% |
| Enzyme ọrụ | ≥60 Kunitz nkeji/mg |
| Isi ihe nke isoelectric | 9.6 |
Ihe ndetu
Maka nchekwa na ahụike gị, biko yiri uwe lab na gloves enwere ike iji rụọ ọrụ.
