Rnase A
Ko te Ribonuclease A (RNaseA) he polypeptide kotahi-waa kei roto e 4 nga here disulfide me te taumaha ngota o te 13.7 kDa.Ko te RNase A he endoribonuclease e tino whakaheke ana i te RNA whenu kotahi ki nga toenga C me U.Ko te tikanga, ka mohio te wehenga ki te here phosphodiester i hangaia e te 5'-ribose o te nucleotide me te roopu phosphate i runga i te 3'-ribose o te pirimidine nucleotide e tata ana, na reira ka hydrolyzed te 2, 3'-Cyclic phosphates ki te 3 e rite ana. ′nucleoside phosphates (hei tauira, pG-pG-pC-pA-pG ka wehea e RNase A hei whakaputa pG-pG-pCp me A-PG).Ko te RNase A te mea tino kaha ki te wawahi i te RNA whenu kotahi.Ko te kukū mahi e manakohia ana ko te 1-100μG/mL, he hototahi ki nga momo punaha tauhohenga.Ka taea te whakamahi i te kukū iti o te tote (0-100 mM NaCl) ki te tapahi i te RNA tautahi, te RNA taurua, me nga mekameka RNA i hangaia e te RNA-DNA hybridization.Heoi, i te nui o te tote (≥0.3 M), ko te RNase A anake te wehe i te RNA pae kotahi.
Ko te RNase A te nuinga o te whakamahi ki te tango i te RNA i te wa e whakarite ana te DNA plasmid, te DNA genomic ranei. Ahakoa kei te kaha te DNase i te wa o te whakaritenga ka taea te pa ki te tauhohenga.Ko te tikanga tuku iho mo te koropupu i roto i te pati wai ka taea te whakamahi hei whakakore i te mahi DNase.Ko tenei hua kaore he DNase me te protease, kaore e hiahiatia he maimoatanga wera i mua i te whakamahinga.I tua atu, ka taea hoki te whakamahi i tenei hua i roto i nga whakamatautau koiora ngota penei i te tātari tiaki RNase me te tātari raupapa RNA.
Nga Tikanga Rokiroki
Ka taea te rongoa i te hua ki te -25～-15℃, whai mana mo te 2 tau.
Tohutohu
Koinei tetahi o nga tikanga noa mo te whakarite RNase A otinga rokiroki.Ka taea hoki te whakarite eetahi atu tikanga e ai ki nga tikanga tuku iho i roto i te taiwhanga, tuhinga tohutoro ranei (pēnei irewa tika i roto i te 10 mM Tris-HCl, pH 7.5, Tris-NaCl wairewa ranei)
1. Whakamahia te 10 mM konutai acetate (pH 5.2) ki te whakarite 10 mg/mL o RNase A otinga rokiroki
2. Whakawera i te 100 ℃ mo te 15 min
3. Matao ki te pāmahana rūma, tāpirihia te 1/10 rōrahi o te 1 M Tris-HCl (pH 7.4), whakatikahia tōna pH ki te 7.4 (motauira, tāpirihia te 500 ml o 10g/ml RNase otinga rokiroki 1M Tris-HCL, PH7.4)
4. Kua whakaemihia ki te -20 ℃ mo te rokiroki tio, ka taea te noho pumau mo te 2 tau.
[Notes]: Ina kohuatia te otinga RNaseA i raro i nga tikanga kore, ka puta te rerenga RNase;Kohuatia ki te iti ake o te pH, a, ki te mea he ua, ka taea te kite, tera pea na te noho o te poke o te pūmua.Mena ka kitea te parataiao i muri i te kohua, ka taea te tango i nga poke ma te tere o te centrifugation (13000rpm), katahi ka whakakiia mo te rokiroki mīti.
Nga korero hua
| Nga kupu taurite | Ribonuclease I;ribonuclease pancreatic;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I |
| CAS No. | 9001-99-4 |
| Te ahua | Te paura lyophilized ma |
| Te taumaha ngota | ~ 13.7kDa (raupapa waikawa amino) |
| Uara Ph | 7.6 (awhe mahi 6-10) |
| Te Pamahana Tika | 60℃ (Awhe ngohe 15-70℃) |
| Kaiwhakahohe | Na2+.K+ |
| Kaitautoko | Rnase Inhibitor |
| Tikanga whakakore | Kaore e taea te whakakore ma te whakamahana, e whakaarohia ana kia whakamahia te pou centrifuge |
| Taketake | Bovine |
| Wairewa | Wairewa ki te wai (10mg/ml) |
| Ngaronga i te maroke | ≤5.0% |
| Te ngohe ngohe | ≥60 wae Kunitz/mg |
| Ikohikohiko | 9.6 |
Notes
Mo to haumaru me to hauora, mau koti taiwhanga me nga karapu tuku mo te mahi.
