Rnase A
Ribonuclease A (RNaseA) waa polypeptide-ka-xidhan oo ka kooban 4 curaarta disulfide oo leh miisaan kelli ah oo ku saabsan 13.7 kDa.RNase A waa endoribonuclease kaas oo si gaar ah hoos u dhigaya RNA hal-xadhigleed ee C iyo U hadhaaga.Gaar ahaan, kala-goynta waxay aqoonsanaysaa dammaanadda fosfodiester-ka ee ay sameeyeen 5'-ribose ee nucleotide iyo kooxda fosfateetka ee 3'-ribose ee nucleotide pyrimidine ee ku xiga, si 2, 3'-fosfatiyeyaasha cyclic lagu nadiifiyo 3 u dhiganta. Fosfooraska nucleoside (tusaale pG-pG-pC-pA-pG waxaa kala jeexjeexay RNase A si ay u dhaliso pG-pG-pCp iyo A-PG).RNase A waa kan ugu firfircoon ee kala jeexjeexa RNA-geesoodka ah.U fiirsashada shaqada ee lagu taliyay waa 1-100μG/ml, oo la jaan qaadaya hababka falcelinta ee kala duwan.Isku-duubnaanta milixdu yar tahay (0-100 mM NaCl) ayaa loo isticmaali karaa in lagu gooyo RNA hal-xadhigle ah, RNA laba-geesood ah, iyo RNA silsilado ay sameeyeen isku-dhafka RNA-DNA.Si kastaba ha ahaatee, marka la eego milix badan (≥0.3 M), RNase A kaliya waxay si gaar ah u jebisaa RNA hal-xadhigle ah.
RNase A waxaa inta badan loo isticmaalaa in laga saaro RNA inta lagu guda jiro diyaarinta plasmid DNA ama genomic DNA. Haddii ay DNA-da firfircoon iyo in kale inta lagu guda jiro habka diyaarinta waxay si fudud u saameyn kartaa falcelinta.Habka dhaqameed ee lagu kariyo biyaha qubeyska ayaa loo isticmaali karaa in la joojiyo dhaqdhaqaaqa DNA-ga.Alaabtani kuma jiraan DNA-ga iyo borotiinka, umana baahna daawaynta kulaylka ka hor inta aan la isticmaalin.Intaa waxaa dheer, badeecadan sidoo kale waxaa loo isticmaali karaa tijaabooyinka bayoloji molecular sida falanqaynta ilaalinta RNase iyo falanqaynta taxanaha RNA.
Xaaladaha Kaydinta
Alaabta waxaa lagu kaydin karaa at -25～-15℃, shaqaynaysa 2 sano.
Tilmaamaha
Tani waa mid ka mid ah hababka caadiga ah ee diyaarinta RNase A xal kaydinta.Waxa kale oo lagu diyaarin karaahababka kale ee waafaqsan hababka dhaqanka ee shaybaarka ama suugaanta tixraaca (sidasi toos ah ugu milma 10 mM Tris-HCl, pH 7.5 ama Tris-NaCl
1. Isticmaal 10 mM sodium acetate (pH 5.2) si aad u diyaariso 10 mg/mL ee RNase A xal kaydinta
2. Kuleylinta 100 ℃ 15 daqiiqo
3. Ku qabow heerkulka qolka, ku dar 1/10 mug 1 M Tris-HCl (pH 7.4), ku hagaaji pH ilaa 7.4tusaale ahaan, ku dar 500 ml oo ah 10g/ml RNase xal kaydinta 1M Tris-HCL, PH7.4)
4. Baakad hoose oo ah -20℃ kaydinta barafaysan, taas oo noqon karta mid xasilloon ilaa 2 sano.
[Xusuusin]: Marka karkarinta RNaseA xalalka dhexdhexaadka ah, roobab RNase ayaa sameyn doona;Ku kari pH hoose, iyo haddii ay jirto roobab, waa la arki karaa, taas oo ay sababi karto joogitaanka borotiinka wasakhaysan.Haddii wasakhda la helo ka dib karkarinta, wasakhda waxaa lagu saari karaa xawaaraha sare ee centrifugation (13000rpm), ka dibna si hoos loogu baakadeeyo kaydinta qaboojinta.
macluumaadka alaabta
| La mid ah | Ribonuclease I;Ribonuclease ee ganaca;Ribonuclease 3'-pyrimidnooligonucleotidohydrolase;Rnase A;Endoribonulcease I |
| CAS Maya. | 9001-99-4 |
| Muuqashada | Budada lyophilized cad |
| Miisaanka molecular | ~ 13.7kDa (isku xigxiga amino acid) |
| Ph Qiimaha | 7.6 (kala duwanaanshaha dhaqdhaqaaqa 6-10) |
| Heerkulka ku habboon | 60℃ (Dhaqdhaqaaqa kala duwan 15-70℃) |
| Wakiil firfircoon | Na2+.K+ |
| Kahortagga | Rnase Inhibitor |
| Habka hawlgelinta | Laguma dhaqaajin karo kuleyliyaha, waxaa la soo jeediyay in la isticmaalo tiirka centrifuge |
| Asal ahaan | Bovine |
| Solubility | Biyo ku milmay (10mg/ml) |
| Khasaare qalalan | ≤5.0% |
| Dhaqdhaqaaqa enzyme | ≥60 Kunitz unug/mg |
| Barta koronto | 9.6 |
Xusuusin
Badbaadadaada iyo caafimaadkaaga, fadlan xidho jaakadaha shaybaadhka iyo galoofyada la tuuri karo ee shaqada.
