High-level Expression and Purification of Untagged and Histidine-tagged Human Immunodeficiency Virus type-1 (HIV-1) Reverse Transcriptase
Description of Invention:
This invention includes plasmids and protocols to express and purify large quantities of histidine-tagged and untagged HIV-1 reverse transcriptase (RT). Conditions have been optimized for overexpression and purification of p66 and p51 heterodimer RT in E. coli. High-level of expression was reached as RT represented approximately 30%-40% of total cell proteins. The subject invention enables the purification of large quantities of heterodimer RT necessary for structural and kinetic studies and facilitates subunit-specific amino acid alterations essential for structure/function investigations.
Application:
Research Tool
Development Status: In vitro data available
Inventors: Samuel H. Wilson (NIEHS) Rajendra Prasad (NIEHS) Esther W. Hou (NIEHS)
Patent Status:
DHHS Reference No. E-141-2007/0 – Research Tool
Patent protection is not being pursued for this technology.
Relevant Publication: EW Hou, R Prasad, WA Beard, SH Wilson. High-level expression and purification of untagged and histidine-tagged HIV-1 reverse transcriptase. Protein Expr Purif. 2004 Mar;34(1):75-86. [PubMed abs]
Licensing Status: Available for non-exclusive licensing as biological material and research tool.
For Additional Information Please Contact: Sally Hu PhD MBA
NIH Office of Technology Transfer
6011 Executive Blvd, Suite 325
Rockville, MD 20852-3804
Phone: (301) 435-5606
Email: hus@mail.nih.gov
Fax: (301) 402-0220
