Ultra Nuclease
UltraNuclease ndi ma genetic engineeredendonuclaese omwe amachokera ku Serratia marcescens, omwe amatha kunyozetsa DNA kapena RNA, kaya yawiri kapena imodzi, yozungulira kapena yozungulira pansi pamitundu yambiri, imasokoneza kotheratu ma nucleic acid kukhala 5'-monophosphate oligonucleotides ndi 3-5base kutalika ndi 3-5. .Pambuyo pa kusinthidwa kwa uinjiniya wa majini, mankhwalawa adayatsidwa, kufotokozedwa, ndikuyeretsedwa mu Escherichia coli (E. coli), yomwe imachepetsa kukhuthala kwa cell supernatant ndi cell lysate kafukufuku wasayansi, komanso kupititsa patsogolo kuyeretsedwa bwino ndi kafukufuku wamapuloteni.Itha kugwiritsidwanso ntchito pamankhwala a jini, kuyeretsa ma virus, kupanga katemera, makampani opanga mankhwala a protein ndi polysaccharide ngati zotsalira za nucleic acid kuchotsa reagent.
Zogulitsa
| CAS No. | 9025-65-4 |
| EC No. | |
| Kulemera kwa Maselo | 30k ndi |
| Isoelectric Point | 6.85 |
| Mapuloteni Purity | ≥99% (SDS-PAGE & SEC-HPLC) |
| Ntchito Yeniyeni | ≥1.1 × 106 U/mg |
| OptimumTemperature | 37°C |
| Mulingo wabwino kwambiri wa pH | 8.0 |
| Ntchito ya Protease | zoipa |
| Bioburden | ＜10CFU/100,000U |
| Residual Host-cell Protein | ≤10ppm |
| Chitsulo Cholemera | ≤10ppm |
| Bakiteriya Endotoxin | ＜0.25EU/1000U |
| Kusungirako Buffer | 20mM Tris-HCl, pH 8.0, 2mM MgCl2, 20mM NaCl, 50% Glycerol |
Zosungirako
≤0 ° C zoyendera; -25~-15 °C Kusungirako, zaka 2 zovomerezeka (peŵani kuzizira-kusungunuka).
Tanthauzo la Chigawo
Kuchuluka kwa enzyme yomwe imagwiritsidwa ntchito posintha mayamwidwe a △A260 ndi 1.0 mkati mwa 30min pa 37 °C, pH 8.0, yofanana ndi 37μg ya umuna wa salimoni wopukutidwa mwa kudula kukhala oligonucleotides, adatanthauzidwa ngati unit yogwira ntchito(U).
Kuwongolera khalidwe
Residual Host-cell Protein: ELISA zida
•Protease Zotsalira: 250KU/mL UltraNuclease adachita ndi gawo lapansi kwa 60min, palibe ntchito yomwe idapezeka.
•Bakiteriya Endotoxin: LAL-Test, Pharmacopoeia of the People's Republic of China Volume 4 (2020 Edition) njira yoyesera malire a gel.General Malamulo (1143).
•Bioburden: Pharmacopoeia of the People's Republic of China Volume 4 (2020 Edition)— General
Malamulo a Kusabereka Mayeso (1101), PRC National Standard, GB 4789.2-2016.
•Chitsulo Cholemera:ICP-AES, HJ776-2015.
Ntchito
Ntchito ya UltraNuclease idalephereka kwambiri pamene kukhazikika kwa SDS kunali kopitilira 0.1% kapena EDTA
kukhazikika kunali kopitilira 1mM. Surfactant Triton X- 100, Tween 20 ndi Tween 80 zinalibe mphamvu pa nuclease
katundu pamene ndende anali pansi 1.5%.
| Ntchito | Mulingo woyenera ntchito | Ntchito Yovomerezeka |
| Kutentha | 37 ℃ | 0-45 ℃ |
| pH | 8.0-9.2 | 6.0-11.0 |
| Mg2+ | 1-2 mm | 1-15 mm |
| Mtengo wa DTT | 0-100 mm | > 100mM |
| 2-Mercaptoethanol | 0-100 mm | > 100mM |
| Monovalent zitsulo ion (Na+, K+ etc.) | 0-20 mm | 0-200 mm |
| PO43- | 0-10 mm | 0-100 mm |
Kagwiritsidwe ndi Mlingo
• Chotsani exogenous nucleic acid ku mankhwala a katemera, kuchepetsa chiopsezo cha nucleic acid yotsalira kawopsedwe ndi kukonza chitetezo cha mankhwala.
• Chepetsani kukhuthala kwa madzi a chakudya chifukwa cha nucleic acid, kufupikitsa nthawi yokonza ndi kuonjezera zokolola.
• Chotsani nucleic acid yomwe idakulungidwa (virus, inclusion body, etc.), yomwe ili yabwino.
kumasulidwa ndi kuyeretsedwa kwa tinthu.
| Mtundu Woyesera | Kupanga Mapuloteni | Virus, Vaccine | Mankhwala Osokoneza Bongo |
| Nambala ya Maselo | 1 g selo yonyowa kulemera (yoyimitsidwanso ndi 10ml buffer) | 1L kuthirira madzi supernatant | 1L chikhalidwe |
| Ochepa Mlingo | 250 U | 100 U | 100 U |
| Analimbikitsa Mlingo | 2500 U | 25000U | 5000 U |
• Chithandizo cha nyukiliya chikhoza kupititsa patsogolo kuthetsa ndi kubwezeretsanso chitsanzo cha column chromatography, electrophoresis ndi blotting analysis.
• Mu chithandizo cha majini, nucleic acid imachotsedwa kuti ipeze mavairasi oyeretsedwa a adeno.
