PAN-1 TAP pEG(KT) plasmid
JHU REF: [C11959]
Invention novelty: This invention is a tangible material in the form of the PAN-1 TAP pEG(KT) plasmid.
Value Proposition: the PAN-1 TAP pEG(KT) plasmid
Technical Details:
This technology provides a tangible material in the form of a PAN-1 TAP-expressing pEG(KT) plasmid, which is an application of the pEG(KT) system for protein expression and purification.
pEG(KT) is a widely used plasmid for expressing high levels of GST fusion proteins in the yeast in a controlled manner. In particular, it imparts a high copy number in yeast cells. Its galactose promoter controls the expression of the desired protein product, which also contributes to high levels of expression when the cells are grown in galactose-containing medium. In addition, it has the leu2-d allele of LEU2, which allows for a further selection of cells that have extremely high numbers of plasmids by growing the cells in medium lacking leucine. The complete sequence of the pEG(KT) plasmid was reported in 2009, further facilitating its applications.
BJ2168 is a protease-deficient yeast strain. The yeast contains a large number of proteases that are located in various compartments and membranes of the cell. Some of the proteases pose significant impediments to analysis of biochemical processes and/or purification of proteins, and can generate artifacts concerning the activity, structure, and, even, intracellular location of proteins. Therefore, the BJ2168 yeast strain can maintain high levels of intact protein expression by removing proteases.
As an example of the application of pEG(KT) expression system, Pan1-TAP-expressing pEG(KT) plasmid was generated and transformed into the BJ2168 yeast strain. High levels of Pan1-TAP protein were produced when the cell were cultured in inducing conditions. This plasmid can be used as a positive control for the pEG(KT) expression system.
Looking for Partners: To license these materials to biotech and/or pharmaceutical companies.
Stage of Development: Tangible Materials
Data Availability:
The complete sequence of the pEG(KT) plasmid has been reported (Pierce et al., 2009).
Pan1-TAP-expressiong pEG(KT) plasmid was generated and transformed into the protease-deficient yeast strain BJ2168. High levels of Pan1-TAP protein were produced when the cell were cultured in inducing conditions.
Patent Status: None – Tangible Material
Publication(s)/Associated Cases:
Pierce BD, Wendland B. Sequence of the yeast protein expression plasmid pEG(KT). Yeast. 2009 Jun;26(6):349-53.
Categories: Research Tools, Tangible Property (plasmids and a yeast strain)
Keywords: pEG(KT) plasmid, PAN-1 TAP pEG(KT), and BJ2168 yeast strain
