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프로모터 : 상시발현 프로모터 (Pichia pastoris TEF promoter) 활용
분비서열 : 단백질 분비를 위한 Signal Peptide 서열
숙주세포 : 야생형 균주 게놈 편집을 통한 단백질 고효율 발현용 균주를 이용
*PpTEF promoter, Pichia pastoris translational elongation factor EF-1 alpha promoter.
Target genome site: bases 18-683
PpTEF* promoter: bases 712-1148
Codon-optimized MR(KR): bases 1149-1403
SapI: bases 1404-1410, 1463-1469
3’-UTR: bases 1474-1547
AOX1 terminator: bases 1548-1794
FRT (minimal): bases 1796-1829
TEF1 Promoter: bases 1847-2244
EM7 promoter: bases 2251-2298
Selection marker: bases 2317-2691
PpGAPDH terminator: bases 2698-3190
pBR322 origin: bases 3199-3787
FRT (minimal): bases 1796-1829, 3794-3827
Type II 제한효소 (Sap I) 사용으로 Target gene Assembly 가능
Target genome site를 적용한 Homologous recombination 가능
TEF-EM7 Hybrid promoter 도입으로 E. coli 와 Yeast (P. pastoris)에서 동일 selection marker (e.g. BleoR) 적용가능
APExpress™ Pichia Expression vector는 재조합 단백질을 yeast Pichia pastoris에서 높은 수준으로 발현 가능
APExpress™ Pichia Expression Kit는 PpTEF promoter에서 목적 유전자를 발현시키기 위해 필수적인 구성품을 제공
Promotor : Glucose regulated constitutive TEF promoter(Pichia pastoris)
Signalpeptide : P. pastoris mating factor α (MFα) for secreted expression from Saccharomyces cerevisiae.
Host strain : Genome editing of wild-type strains for high-efficiency protein expression
*PpTEF promoter, Pichia pastoris translational elongation factor EF-1 alpha promoter.
Target genome site: bases 18-683
PpTEF* promoter: bases 712-1148
Codon-optimized MR(KR): bases 1149-1403
SapI: bases 1404-1410, 1463-1469
3’-UTR: bases 1474-1547
AOX1 terminator: bases 1548-1794
FRT (minimal): bases 1796-1829
TEF1 Promoter: bases 1847-2244
EM7 promoter: bases 2251-2298
Selection marker: bases 2317-2691
PpGAPDH terminator: bases 2698-3190
pBR322 origin: bases 3199-3787
FRT (minimal): bases 1796-1829, 3794-3827
Type II restriction enzyme(SapI) in the multiple cloning site for target gene assembly
TEF-EM7 Hybrid promoter for select-able marker(e. g. BleoR) application both E. coli and yeast (P. pastoris).
APExpress™ pichia Expression kit is manufactured for high yield recombinant protein expression in yeast Pichia pastoris
APExpress™ pichia Expression kit is supplied all component for high level expression of recombinant proteins
The AOX1 promoter is used to drive expression of the gene of interest encoding the desired heterologous protein in APExpress™ pichia Expression System.