Invitrogen™

Champion™ pET200 Directional TOPO™ Expression Kit with BL21 Star™ (DE3) One Shot™ Chemically Competent E. coli

The Champion™ pET Expression System yields the highest-level protein production in E. coli. During expression, your protein of interest can자세히 알아보기

The Champion™ pET Expression System yields the highest-level protein production in E. coli. During expression, your protein of interest can reach levels greater than 50 percent of total cellular protein. Based on T7 expression vectors originally developed by Studier and colleagues (1-3), high-level expression is achieved because the T7 RNA polymerase is more processive than native E. coli_RNA polymerase and is dedicated to the transcription of your gene of interest. Protein production is further enhanced in the system by the expression strain BL21 Star™ _E. coli, which significantly improves the stability of mRNA transcripts and increases protein expression up to ten-fold.

Simplified, efficient Directional TOPO™ cloning allows you to quickly enter a Champion™ pET Expression vector. These kits feature linearized, topoisomerase I-activated Champion™ pET expression vectors for five-minute directional cloning. Directional TOPO™ Cloning technology facilitates gene expression because:

• A proofreading enzyme is used for PCR, resulting in fewer errors in cloned genes
• Greater than 90% of the clones are in the correct orientation for gene expression, reducing the time spent on colony screening

Seven Champion™ pET Directional TOPO™ Expression Vectors are available (Figure 1 and Table 1): Each vector carries a T7_lac_ promoter for high-level expression. Flexible options for simplifying protein detection, cleaving purification tags, selecting plasmid carrying clones, and/or improving protein yields are available.
With the Champion™ pET Directional TOPO™ vectors you can expect highest-level protein production. Figure 2 shows expression of the _lac_Z gene in Champion™ pET Directional TOPO™ vectors. Figure 3 demonstrates efficient cleavage using TEV protease of the N-terminal tag of a β-galactosidase fusion protein expressed from pET151/D-TOPO™.

사양

구성 또는 유도성 시스템Inducible

유도제IPTG

프로모터T7, lacO

제품 유형TOPO Expression Kit

선택 제제(진핵)None

항생제 내성 박테리아Kanamycin (KanR)

박테리아 또는 효모 균주BL21 Star™(DE3)

분열EK (Enterokinase) Recognition Site

발현 시스템E. coli

발현 메커니즘Cell-Based Expression

단백질 태그His Tag (6x), Xpress Epitope Tag

클로닝 방법Directional TOPO

수량20 reactions

벡터pET

제품라인One Shot™

Unit SizeEach