e707004304
ORANGE EKSTRAKLASA
Dołączył: 17 Gru 2010
Posty: 612
Przeczytał: 0 tematów
Ostrzeżeń: 0/5
Skąd: England
|
 Wysłany: Czw 20:50, 07 Kwi 2011 |
|
|
A rapid screening Pichia pastoris expression clone of the new method of high
astor / sclon ~ [J]. Biotech. nology, 1996,2 O (6) :980-982 .1 i1J1J] J716 of Jinan University (Medical Sciences ) 20O4 years [7] [8] SCORERCA, CLAREJJ, MOCOMBINWR, eta1. RapidselectionusingG418ofhighcopynumbertran ~ ormantsofP / ch / apastor / sforhigh-levelforeigngeneexpression [J]. Biotechnology, 1994,12 (2) :181-184. JEFFREYTM, DANL, BRADD, eta1. ExpressionoftrimericCD40ligandinP/ch/apastor/s: useofarapidmethodtodetecthigh-levelexpressingtransformants [J]. Gene ,1997,187:193-200. Anewmethodforrapidscreenofhigh--expressingP/ch/apastor/sclonesXIEQi-xuan, XIAOLuan-juan,[link widoczny dla zalogowanych], PANShah-pei, YUWei, PENGYa-lin, ZHANGChun-xue (CenterforReproductiveImmunologyResearch, JinanUniversity, Guangzhou510632, China) lAbstractjAim_, Forscreeninghigh-expressingrecombinantPichiapastorisclonesexactlyandquickly. Methods: AccordingtophysiologicalcharacteristicofPichiapastorisandthespecialtiesofEuSA-Dot, asimplemethodwasdevelopedtoidentifythehigh-expressingtransformantsfromlargeaxnotmtsofclonesinthes {u11etime. GS115 a thZP3recombinantswereinoeulatodonaMMplatetoformclones, thenaNCmembranewaslaidontopoftheplate. Mterwettedbythesecretiveproduces, theNCmembraneWasremovedanddetectedbytheEUsA-DotwiththepolyantibodiestoporcineZP3. Thehigh-expressingcloneswereselectedtoproducetherhintheshakeflasksandthesupemateswereiden a edbytheSDS-PAGEandWesternBlotting. Results: eresultinEUsA-DotWasconfmnedbytheSDS-PAGEandtheWesternBlotting. Conclusion: ThemethodiSmorehigh-flux. quickandeasythanthetraditionalmethodandcallbeusedtoscreentheengineeringPichiapastorisclones. [Keywords] Pichiapastoris; clone; screen; ELISA-Dot [ Editor: Huang Jianjun ]
Post został pochwalony 0 razy
|
|
