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1、酵母基因克隆与表达载体（Gene cloning and expression vector of yeast）Two. Yeast gene cloning and expression vector(a). Functional skeleton vector - for functional analysis and vector reconstruction for different purposes1, the yeast genome integration plasmid (Yeast integrating plasmid, YIP)(1).Trp (2).Ura scree
2、ning marker selection marker (3).Leu (4).His screening marker selection marker (5).G418 screening markerThis plasmid was linearized to be integrated into the yeast genomePurpose: the integration of A. gene deletion of B. gene substituted or stable expression element (integration) and type (knock in)
3、2, yeast centromeric - autonomously replicating plasmid (Yeast CEN-ARS plasmid, YCP)(1).Trp (2).Ura screening marker selection marker (3).Leu (4).His screening marker selection marker (5).G418 screening markerThis plasmid could self replicate in cells, and due to the presence of centriole replicatio
4、n sequence, so each cell almost only one copy.Purpose: because each cell average only one copy, avoid the expression of target gene in surplus, which is especially suitable for the research of gene function3, yep (Yeast episomal, plasmid, YEP)(1).Trp (2).Ura screening marker selection marker (3).Leu
5、 (4).His screening marker selection marker (5).G418 screening markerThis plasmid replication in cells with highly episomal form, each cell is up to 200-400 a copy of babePurpose: for the expression of exogenous gene(two) expression vector.1, GAL1 promoter expression vector, expression vector induced
6、 by D-galactose(1).Trp (2).Ura screening marker selection marker (3).Leu (4).His screening marker selection marker (5).G418 screening marker2, Cup1 promoter expression vector, induced by copper sulfate(1).Trp (2).Ura screening marker selection marker (3).Leu (4).His screening marker selection marker
7、 (5).G418 screening marker3, ADH promoter expression vector -ADC1 constitutive expression vector(1).Trp (2).Ura screening marker selection marker (3).Leu (4).His screening marker selection marker (5).G418 screening markerYeast genetics research and gene function, higher biological magic (1) expressi
8、on of exogenous gene in yeast2010-07-24 11:51:11| classification: | label method and yeast genetics: yeast culture medium selection defect of dropout expression plasmid | report | name subscriptionAuthor: Pan model Department of biotechnology group Zhu Leinuo KenoBudding yeast (budding yeast, Saccha
9、romyces cerevisiae) commonly known as Saccharomyces cerevisiae, unicellular eukaryotes, its genome is relatively simple (only 3-4 times of Escherichia coli), easy to operate, the genetic background is relatively clear, and the mutant and easy separation, known as eukaryotic Escherichia coli (Eukaryo
10、tic E.coli) and inner tube (In vivo test tube), which is an ideal model organism for studying eukaryotic gene function. Just cannot do without the operation of Escherichia coli, one to the depth development of the molecular biology laboratory cannot do without yeast experimental operation.As the mol
11、ecular biology operation desk data - (Current Protocol in Molecular Biology redpaper) and column yeast as a separate section in parallel with other commonly used molecular biology experimental technique; and was regarded as the Bible of the Life Science Laboratory of molecular cloning, multiple sect
12、ions of the third version of the yeast with different aspects of the experimental operation, which shows an important role in todays yeast genetics method in life science research. Yeast in life science research is mainly used in the following aspects:Foreign gene expression in yeast (Foreign gene e
13、xpression in the yeast)However, most commonly used exogenous gene expression receptor is Escherichia coli, and there are great differences between codon preference between the nature of some species, some species of the gene can be effectively expressed in Escherichia coli, and some species of the g
14、enes expressed in Escherichia coli is very weak or unable to express (such as some forest plants). In this case, as eukaryotic yeast gene expression has become the first choice of receptor cells. Pan Keno company technical department successfully for forestry scientific research units expressed a lo
15、t of protein with natural activity, and these proteins can not be expressed in prokaryotic Escherichia coli (the theory and technology of more detailed information can be emailed to: fungenome consulting). In addition, the yeast is easy to breed characteristics and non-toxic (daily life used to fer
16、ment flour and wine, it is also known as the bread yeast and beer yeast), the biological technology and industry researchers used yeast to exogenous gene expression, get a lot of protein with natural activity and production of genetically engineered vaccine.The choice of the expression vector of 1. yeastUsing yeast to obtain high yield protein, its expression vector
