Thermo Fisher K10001 20 reactions Champion pET100Directional TOPOExpression kit, including BL21 Star (DE3) One ShotChemical Competence<i>Escherichia coli</i>
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K10001 20 reactionsChampion pET100 Targeted TOPO Expression Kit, containing BL21 Star (DE3) One Shot Chemically Competent E. coli

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Champion™ pET Expression System produces the highest levels of protein yields in E. coli. During expression, the protein of interest may reach levels exceeding 50% of total cellular protein. Based on the T7 expression vector originally developed by Studier and colleagues (1-3), because T7 RNA polymerase is more synthetic than the natural E. coli RNA polymerase and is dedicated to the transcription of the gene of interest, this can be achieved High level expression. Expression strain BL21 Star ™E. coli can further improve the protein yield in the system, significantly improve the stability of mRNA transcripts and increase protein expression tenfold.

Simplified and efficient directional TOPO™ cloning provides rapid access to Champion™ pET expression vectors. These kits feature linearized, topoisomerase I-activated Champion™ pET expression vectors for 5-minute directional cloning. Directional TOPO™ cloning technology promotes gene expression because:

•Proofreading enzymes are used in PCR, resulting in fewer errors in cloned genes
•More than 90% of clones achieve the correct direction of gene expression , thereby reducing the time spent on colony screening

Seven Champion™ pET-directed TOPO™ expression vectors are available (Figure 1 and Table 1): Each vector carries a T7lac Promoter for high-level expression. Offers flexible options to simplify protein detection, cleave purification tags, select plasmid-bearing clones, and/or increase protein yields.
Achieve the highest levels of protein yield with Champion™ pET oriented TOPO™ vectors. Figure 2 shows the expression of thelacZ gene in the Champion™ pET-directed TOPO™ vector. Figure 3 shows efficient cleavage of the N-terminal tag of the β-galactosidase fusion protein expressed from pET151/D-TOPO™ using TEV protease.
For Research Use Only. Not for use in diagnostic procedures.