High-level Expression and Purification of Untagged and Histidine-tagged Human Immunodeficiency Virus type-1 (HIV-1) Reverse Tran
Posted Jun 13 2010 5:00pm
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.
Applications: Research Tool
Development Status: In vitro data available
Inventors: Samuel H Wilson (NIEHS) Rajendra Prasad (NIEHS) Ester W Hou (NIEHS)
Patent Status: 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.
Portfolios: Infectious Diseases Infectious Diseases - Research Materials Infectious Diseases - Other In-vitro Data
For Additional Information Please Contact: Sally Hu Ph.D., M.B.A. NIH Office of Technology Transfer 6011 Executive Blvd. Suite 325
Room 21, Rockville, MD 20852 United States Email: hus@mail.nih.gov Phone: 301-435-5606 Fax: 301-402-0220
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.
Applications:
Research Tool
Development Status:
In vitro data available
Inventors:
Samuel H Wilson (NIEHS)
Rajendra Prasad (NIEHS)
Ester W Hou (NIEHS)
Patent Status:
Research Tool -- patent protection is not being pursued for this technology
Relevant Publication:
Licensing Status:
Available for non-exclusive licensing as biological material and research tool.
Portfolios:
Infectious Diseases
Infectious Diseases - Research Materials
Infectious Diseases - Other
In-vitro Data
For Additional Information Please Contact:
Sally Hu Ph.D., M.B.A.
NIH Office of Technology Transfer
6011 Executive Blvd. Suite 325 Room 21,
Rockville, MD 20852
United States
Email: hus@mail.nih.gov
Phone: 301-435-5606
Fax: 301-402-0220
Ref No: 1517
Updated: 06/2010