Protein expression vectors for commercial use

Takara Bio's pCold Expression Vectors offer cold shock expression technology for high purity, high yield protein production. The series includes four different vectors that utilize the cold shock Protein A (cspA) promoter to direct expression of recombinant protein in E. coli. These vectors selectively induce target protein synthesis at low temperatures (15°C) when the synthesis of other proteins is suppressed and protease activity is decreased. This results in high yield of the target protein (60% of intracellular protein). In addition to the cspA promoter, all four vectors (including pCold I) contain a lac operator (for control of expression), ampicillin resistance gene (amp^r), ColE1 origin of replication, M13 IG fragment, and multiple cloning site (MCS). pCold I also contains a Factor Xa cleavage site. These vectors work equally well for synthesis of non-labeled and radiolabeled proteins and can be used in conjunction with Takara Bio's Chaperone Plasmid Set (Cat.# 3340P).

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3361: pCold I DNA

Takara's pCold Expression Vectors offer cold shock expression technology for high purity, high yield protein production. The series includes four different vectors that utilize the cold shock Protein A (cspA) promoter to direct expression of recombinant protein in E. coli. These vectors selectively induce target protein synthesis at low temperatures (15°C) when the synthesis of other proteins is suppressed and protease activity is decreased. This results in high yield of the target protein (60% of intracellular protein). In addition to the cspA promoter, all four vectors contain a lac operator (for control of expression), ampicillin resistance gene (amp^r), ColE1 origin of replication, M13 IG fragment, and multiple cloning site (MCS). Furthermore, pCold III DNA also contains a his tag sequence and a Factor Xa cleavage site. These vectors work equally well for synthesis of non-labeled and radiolabeled proteins and can be used in conjunction with Takara's Chaperone Plasmid Set (Cat.# 3340P).

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3363: pCold III DNA

The pCold TF DNA Vector provides cold shock technology for high yield protein expression combined with Trigger Factor (chaperone) expression to facilitate correct protein folding, thus enabling efficient soluble protein production for otherwise intractable target proteins. pCold TF DNA Vector is a fusion cold shock expression vector that expresses Trigger Factor (TF) chaperone as a soluble tag. Trigger Factor is a prokaryotic ribosome-associated chaperone protein (48 kDa) which facilitates co-translational folding of newly expressed polypeptides. Because of its E. coli origin, TF is highly expressed in E. coli expression systems. The pCold TF DNA Vector consists of the cspA promoter plus additional downstream sequences including a 5' untranslated region (5' UTR), a translation enhancing element (TEE), a His-Tag sequence, and a multicloning site (MCS). A lac operator is inserted downstream of the cspA promoter to ensure strict regulation of expression. Additionally, recognition sites for HRV 3C Protease, Thrombin, and Factor Xa are located between TF-Tag and the Multiple Cloning Site (MCS) and function to facilitate tag removal from the expressed fusion protein. E. coli strains can serve as expression hosts.

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3365: pCold TF DNA

The Chaperone Plasmid Set consists of five different plasmids, each of which is designed to express multiple molecular chaperones that function as a "chaperone team" to enable protein folding. Co-expression of a target protein with one of these chaperone teams increases the recovery of soluble proteins. Each plasmid carries an origin of replication derived from pACYC and a Cm^r gene, which allows use with E. coli expression systems utilizing ColE1-type plasmids containing an ampicillin resistance gene as a marker. The chaperone genes are situated downstream of an araB or Pzt-1 (tet) promoter. Therefore, expression of target proteins and chaperones can be induced individually if the target gene is placed under the control of other promoters (e.g. lac). These plasmids also contain the necessary regulator (araC or tet^r) for each promoter.

Use this set to perform a two step method to construct target/chaperone co-expression systems: In the first step prepare an E. coli host transformed with only the chaperone plasmid. The second step is to prepare competent cells from this new strain and transform this strain with a plasmid expressing the target protein.

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3340: Chaperone Plasmid Set