Purification of Chlamydia Antigen Pgp3

The selection and isolation of the CT antigen Pgp3 has been previously described.^{6 (link)} Briefly, XL1 Blue bacterial cultures containing Pgp3 (Pgp3 cloned from serovar E) in a pGEX6 (Amersham Biosciences, Piscataway, NJ) vector system were grown to an optical density of 0.7–0.8, induced with 0.2 MM IPTG, harvested after 2 hours, and the Pgp3-GST fusion protein purified over a glutathione Sepharose 4B affinity column (GE Healthcare, Piscataway, NJ).

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Gwyn S., Cooley G., Goodhew B., Kohlhoff S., Banniettis N., Wiegand R, & Martin D.L. (2017). Comparison of Platforms for Testing Antibody Responses against the Chlamydia trachomatis Antigen Pgp3. The American Journal of Tropical Medicine and Hygiene, 97(6), 1662-1668.

Publication 2017

glutathione Sepharose 4B affinity column

Antigen Bacterial Glutathione Grown Iptg Isolation Protein Sepharose 4b Vector

Corresponding Organization :

Other organizations : The Centers, Centers for Disease Control and Prevention, Division of Parasitic Diseases and Malaria, SUNY Downstate Health Sciences University, State University of New York

Top 5 similar protocols

Variable analysis

independent variables

Induction with 0.2 mM IPTG

dependent variables

Optical density of bacterial cultures
Purification of Pgp3-GST fusion protein

control variables

Growth of XL1 Blue bacterial cultures containing Pgp3 (Pgp3 cloned from serovar E) in a pGEX6 vector system
Harvesting of bacterial cultures after 2 hours
Purification of Pgp3-GST fusion protein over a glutathione Sepharose 4B affinity column

controls

No positive or negative controls were explicitly mentioned in the provided information.

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