Pichia pastoris Expression Kit | expression element library

Pichia pastoris Expression Kit | expression vector + element library

For optimization of various protein expression applications using Pichia pastoris as host cells. Suitable for optimizing the yield of protein scale expression in the laboratory or industrial pilot/scale fermentation.

Please contact us (mail to:support@GMExpression.com) for pre-sales technical consulting and acquire detailed instructions and related DNA sequence files before purchase.

Each element packaged within a 2ml centrifuge tube,  40 µL of 0.5 µg/µL (20 µg circular DNA) in TE buffer, pH 8.0

 The kit including 

Expression vector backbone

Bgl2-pGAP-EcoRI-KZS-α-factor-MSC1(GOI)-ttAOX1+BamHI-PpADE2-arsTRP2

T/X promoters library 

Constitutive   pGAP - pGAPm1 - pGAPm2 - pGAPm6 - p0627

Inducable       pAOX1 - pAOX1m2 - p0472 - pPGK - pADH3 

MCSs library 

MCS1(StuI/KpnI)-MCS2(EcoRI/ BamHI)-MCSres(XbaI/BglII--EcoRI/EcoRV)-MCSgga(BbsI/BsaI--BsaI/BbsI)

T/X terminators library 

ttAOX1 -  ttCYC1

Selection library

ADE2--BleoR(-Zeocin)

ARS library 

arsAOX1 -arsTRP2 -arsHIS4

 Excluding  (purchase separately) 

       Secretion signal library 

α-mating factor pre-sequence + cargo peptide (pro region)

α-mating factor cargo peptide mutant 1

α-mating factor cargo peptide mutant 2

α-amylase signal

Glucoamylase signal

Serum albumin signal

Inulinase signal

Invertase signal

Killer protein signal 

Lysozyme signal

Easy use of the element library

Only 2REs are needed, to achieve 4 GOI’s pick out

Pichia Expression Optimization principles on molecular genetics scale

• More recombination sites (ARS) lead to more copies of GOI.

• High-performance TX/TL elements in the right position and in good order.

• (Not always) Strong (tight regulated /inducible) and suitable TX promoter

• Ribosome binding-site --KZ sequence

• 1~4bp AT-rich UTR

• Suitable TT

• Avoid mRNA hairpin structures

• Low GC regions of large fragments (while high GC large fragments are difficult to synthesize)

• T/L 

• Strong start/ stop codon

• Avoid rare codons by codon optimization

• Pay attention to cytotoxic

• Postmodification

• Glycosylation

• Transport signal

• Matching of key metabolic genes to nutrient/metabolite environments

Note

This item is packaged with 2.8kg of dry ice as standard for international shipping. For domestic shipping, it is packaged with a 1.3kg ice bag.