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Sequence-Validated and Expression-Tested Human cDNA in a Dual Expression,,,Vector

2 consensus sequences permit mRNA to be translated efficiently.

To create the GeneConnection expression-tested clone collection, the gene encoding b-lactamase (ampr) between the Eam1104 I sites in the pDual GC vector is replaced with a human ORF using the seamless cloning method3.

Clone Selection and Availability

GeneConnection expression-tested clones were selected based on their biological function and potential applications. They include kinases, DNA-binding proteins, transferases, transporters, oncogenes, cytochromes, proteases, inflammatory response proteins, cellular matrix proteins, metabolic proteins, synthases, esterases, zinc-finger proteins, and ribosomal proteins. For further information regarding clone selection and availability, visit www.stratagene.com.

Validated Nucleotide Sequence

The nucleotide sequence of the ORF of all expression-tested clones has been determined by double-pass sequencing. A comparison between the clone and a published sequence is provided with each clone.

Epitope and Purification Tags

All clones express a fusion protein consisting of the protein encoded by the cDNA insert, the thrombin cleavage site, three copies of the c-myc epitope tag, and a single copy of the HIS6 purification tag ( Figure 1). The c-myc epitope is derived from the human c-myc gene and contains 10 amino acid residues (EQKLISEEDL).4 This allows for convenient and sensitive detection of expressed proteins with anti-c-myc antibody. The HIS6 purification tag consists of six histidine residues and permits quick and easy purification of the fusion protein from bacterial cells.5

Constitutive High-Level Mammalian Expression


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