pD1204-NHA BioVector NTCC质粒载体菌种细胞基因保藏中心

pD1204-NHA BioVector NTCC质粒载体菌种细胞基因保藏中心 vectors for protein expression in the yeasts Saccharomyces cerevisiae and Pichia pastoris.
Saccharomyces Expression Vectors
These vectors are available with or without secretions signals. They have either an inducible (Gal), strong constitutive (GPD, TEF) or weaker
constitutive (ADH) promoter. The vectors carry LEU2, HIS3, URA3 or TRP1 markers or geneticin resistance. They are maintained on 2μm ori
plasmids.
Expression Protocol
Transform plasmid into appropriate auxotrophic strain, plate onto minimal media lacking the appropriate nutrient and incubate at 30℃ until
heterotrophic colonies arise in 2-3 days. Pick colonies into minimal media or YPD and grow for 24-90 hours at 28-30℃. Check supernatant for
secreted protein expression or cell pellets for cytoplasmic protein expression.
Feature list descriptions
3'UTR_CYC1 The 3'UTR controls post transcriptional regulation. CYC1 is the heme-containing component of the cytochrome b-c1 complex, which accepts
electrons from Rieske protein and transfers electrons to cytochrome c in the mitochondrial respiratory chain. (www.genecards.org/cgi-bin/
carddisp.pl?gene=CYC1 www.genesdev.cshlp.org/content/9/23/2997.full.pdf+html)
3'UTR_URA3 The 3'UTR of the URA3 gene controls post transcriptional regulation. The URA3 gene encodes orotidine-5' phosphate decarboxylase, an enzyme
that is required for the biosynthesis of uracil. (www.genesdev.cshlp.org/content/9/23/2997.full.pdf+html www.protocols.mmml.nl/protocols/yeast/
yeast_selectable_markers.php)
Ampicillin-r A semi-synthetic penicillin derived from 6-amino-penicillanic acid causes cell death by inhibiting cell wall biosynthesis. The gene coding for
ampicillin resistance (bla) is a beta lactamase which is secreted into the periplasmic space where it catalyzes hydrolysis of the beta-lactam ring
of ampicillin. E.coli transformed with plasmid containing the ampicillin resistance gene can grow on media containing 50-100 μg/ml ampicillin.
(www.jac.oxfordjournals.org/content/43/5/699.full)
HA(3) Human influenza haemagglutinin (HA) is a surface glycoprotein required for infectivity of the human virus. The HA tag is derived from the HAmolecule
corresponding to amino acids 98-106 has been extensively used as a general epitope tag in expression vectors. The HA tag does
not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and
purification of the proteins. (www.nature.com/nmeth/journal/v4/n2/full/nmeth0207-107.html www.en.wikipedia.org/wiki/HA-tag)
Ori_2um The 2μ plasmid encodes proteins that allow cells to maintain 20-50 copies of any plasmid carrying the 2μ origin of replication. Because 2μ
plasmids are maintained at such high copy numbers, they provide a convenient way to monitor the effects of overproduction of a particular gene
product. (www.ncbi.nlm.nih.gov/pubmed/2691836)
Ori_pUC The origin of replication is a sequence in a genome at which replication is initiated. The pUC ori is a mutated form of origin derived from E. coli
plasmid pBR322 which allows production of greater than 500 copies of plasmid per cell. (www.en.wikipedia.org/wiki/Origin_of_replication)
P_Gal1 A galactose inducible promoter in Saccharomyces cerevisiae. The GAL1 promoter is repressed by glucose. (www.megrc.re.kr/images/pdf/paper/
paper_in13.pdf www.mcb.asm.org/content/10/9/4757.short)
P_URA3 A promoter from the yeast pyrimidine biosynthetic gene, URA3. (www.ncbi.nlm.nih.gov/pubmed/2204810)
URA3 The URA3 gene encodes orotidine-5' phosphate decarboxylase, an enzyme that is required for the biosynthesis of uracil.
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
电话：+86-010-53513060
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