I-Ultra Nuclease
I-UltraNuclease yi-genetically engineeredendonuclaese ephuma kwi-Serratia marcescens, ekwaziyo ukuthobisa i-DNA okanye i-RNA, nokuba iphindwe kabini okanye i-single stranded, i-linear okanye isetyhula phantsi koluhlu olubanzi lweemeko, ithobisa ngokupheleleyo i-nucleic acids ibe yi-5'-monophosphate oligonucleotides kunye nobude obu-3-5. .Emva kokuguqulwa kobunjineli bemfuza, imveliso iye yabiliswa, yachazwa, yaza yahlanjululwa kwi-Escherichia coli (E. coli), enciphisa i-viscosity ye-cell supernatant kunye ne-cell lysate yophando lwenzululwazi, kodwa iphucula impumelelo yokucoca kunye nokusebenza kophando lweprotheyini.Isenokusetyenziswa kunyango lwemfuza, ukucocwa kwentsholongwane, ukuveliswa kwesitofu sokugonya, iprotein kunye nepolysaccharide kwishishini lamayeza njengentsalela ye-nucleic acid yokususa i-reagent.
Iimpawu zemveliso
| Inombolo yeCAS. | 9025-65-4 |
| Inombolo yeEC. | |
| Ubunzima beMolekyuli | 30kDa |
| Indawo yombane | 6.85 |
| Iprotheyini ecocekileyo | ≥99% (SDS-PAGE & SEC-HPLC) |
| Umsebenzi othile | ≥1.1×106U/mg |
| I-OptimumTemperature | 37°C |
| Eyona pH | 8.0 |
| ProteaseActivity | embi |
| Umthwalo wezinto eziphilayo | ＜10CFU/100,000U |
| Iprotheyini eshiyekileyo yomamkeli-seli | ≤10ppm |
| Intsimbi enzima | ≤10ppm |
| I-Endotoxin yebhaktiriya | ＜0.25EU/1000U |
| Isithinteli sokuGcina | 20mM Tris-HCl, pH 8.0, 2mM MgCl2 , 20mM I-NaCl, i-50% yeGlycerol |
Iimeko zokugcina
≤0 ° C ukuthuthwa; -25 ~ -15 ° C Ukugcinwa, iminyaka eyi-2 esemthethweni (ugweme ukunyibilika kweqhwa).
Inkcazelo yeyunithi
Ubungakanani be-enzyme esetyenziselwa ukutshintsha ixabiso lokufunxa le-△A260 nge-1.0 ngaphakathi kwe-30min kwi-37 °C, i-pH 8.0, ilingana ne-37μg ye-salmon ye-salmon ye-DNA egayiweyo ngokusikwa kwi-oligonucleotides, yachazwa njengeyunithi esebenzayo (U).
Ulawulo lwemeko
Iprotheyini eshiyekileyo yomamkeli-seli: ELISA kit
•Iprotease Iintsalela: I-250KU / mL I-UltraNuclease iphendule nge-substrate ye-60min, akukho msebenzi ufunyenwe.
•I-Endotoxin yebhaktiriya: Uvavanyo lwe-LAL, i-Pharmacopoeia yeRiphabhlikhi yaBantu baseTshayina uMqulu 4 (uHlelo lwe-2020) indlela yokuvavanya umda wejeli.Imigaqo Jikelele (1143).
•Ubunzima bokuphila: I-Pharmacopoeia yeRiphabhlikhi yaBantu baseTshayina uMqulu 4 (uHlelo luka-2020)— Ngokubanzi
Imithetho yoVavanyo lweNzalo (1101), i-PRC yoMgangatho weSizwe, i-GB 4789.2-2016.
•Intsimbi enzima:I-ICP-AES, HJ776-2015.
Ukusebenza
Umsebenzi we-UltraNuclease wawunqandeke kakhulu xa ugxininiso lwe-SDS lungaphezu kwe-0.1% okanye i-EDTA
Ugxininiso lwalungaphezu kwe-1mM.I-Surfactant Triton X- 100, Phakathi kwe-20 kunye ne-Tween 80 yayingenayo impembelelo kwi-nuclease
iipropati xa ugxininiso lungaphantsi kwe-1.5%.
| Ukusebenza | Ukusebenza ngokufanelekileyo | Ukusebenza okusebenzayo |
| Ubushushu | 37℃ | 0-45℃ |
| pH | 8.0-9.2 | 6.0- 11.0 |
| Mg2+ | 1-2mmM | 1- 15mM |
| DTT | 0- 100mM | > 100mM |
| 2-Mercaptoethanol | 0- 100mM | > 100mM |
| Monovalent metal ion (Na+, K+ njl.) | 0-20mmM | 0-200mM |
| PO43- | 0- 10mM | 0- 100mM |
Ukusetyenziswa kunye nomthamo
• Susa i-nucleic acid yangaphandle kwiimveliso zokugonya, yehlisa umngcipheko wentsalela yetyhefu ye-nucleic acid kwaye uphucule ukhuseleko lwemveliso.
• Nciphisa i-viscosity yolwelo lwefidi olubangelwa yi-nucleic acid, nciphisa ixesha lokulungiswa kunye nokwandisa isivuno seprotheyini.
• Susa i-nucleic acid esongiweyo (intsholongwane, umzimba oqukayo, njl.njl.), eluncedo.
ukukhululwa kunye nokuhlanjululwa kweqhekeza.
| Uhlobo loMfuniselo | Ukuveliswa kweeProtheyini | Intsholongwane, isitofu | Iziyobisi zeSeli |
| Inombolo yeeseli | 1g iseli ubunzima emanzi (iphinde yanqunyanyiswa nge-10ml buffer) | 1L ukubiliswa ulwelo olunamandla | 1L inkcubeko |
| Ubuncinci bedosi | 250U | 100U | 100U |
| Umthamo ocetyiswayo | 2500U | 25000U | 5000U |
• Unyango lwe-nuclease lunokuphucula isisombululo kunye nokubuyiswa kwesampulu yekholomu ye-chromatography, i-electrophoresis kunye nohlalutyo lwe-blotting.
• Kwi-gene yonyango, i-nucleic acid iyasuswa ukufumana iintsholongwane ezihlambulukileyo ezihambelana ne-adeno.
