BMB Reports
- Volume 33 Issue 4
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- Pages.337-343
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- 2000
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- 1976-670X(eISSN)
Expression of Human Immunodeficiency Virus Type 1 Tat Proteins in Escherichia coli and Application to Study Tat Functions
- Park, Jin-Seu (Department of Genetic Engineering, Division of Life Sciences, Hallym University) ;
- Lee, Han-Gyu (Department of Genetic Engineering, Division of Life Sciences, Hallym University) ;
- Lee, Yoon (Department of Genetic Engineering, Division of Life Sciences, Hallym University) ;
- Kang, Young-Hee (Department of Food and Nutrition, Division of Life Sciences, Hallym University) ;
- Rhim, Hyang-Shuk (Research Institute of Molecular Genetics, Catholic Research Institute of Medical Science, The Catholic University of Korea) ;
- Choi, Soo-Young (Department of Genetic Engineering, Division of Life Sciences, Hallym University)
- Received : 2000.05.31
- Accepted : 2000.06.29
- Published : 2000.07.31
Abstract
The human immunodeficiency virus type 1 (HIV-1), transactivator of transcription (Tat), is one of the viral gene products that is essential for HIV-1 replication. The HIV-l Tat protein regulates transcription from an HIV-1 long terminal repeat (LTR) and affects the gene expression of cellular proteins during infection. In order to develop an expression system to overexpress and simply purify HIV-1 Tat proteins, the HIV-1 Tat coding sequences that contain one or two exons were amplified using PCR and cloned into a pET vector, which contains a consecutive stretch of six histidine residues at the amino-terminus. The reconstituted vectors were overexpressed in the E. coli strain and the soluble recombinant proteins were purified to be homogeneity in a single step by