We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Expression, purification and characterization of TAT-high mobility group box-1A peptide as a carrier of nucleic acids.
- Authors
Kyunghwa Kim; Jee Seung Han; Hyun Ah Kim; Minhyung Lee
- Abstract
High mobility group box 1 (HMGB1) is an abundant nuclear protein that binds to double-stranded DNA. HMGB1 is composed of high mobility (HMG) box A, box B, and C-terminal acidic regions. In this study, a recombinant TAT linked HMGB1 box A (rTAT-HMGB1A) peptide was expressed, purified, and characterized as a carrier of nucleic acids. The HMGB1A cDNA was amplified by PCR, and cloned into the pET21a expression vector with the TAT domain located at the N-terminus. The rTAT-HMGB1A peptide was overexpressed and purified using Nickel affinity chromatography. A recombinant HMGB1A (rHMGB1A) peptide without the TAT domain was also overexpressed and purified as a control. In gel retardation assays, both the rHMGB1A and rTAT-HMGB1A peptides formed complexes with DNA equally well. However, transfection assays showed that the rTAT-HMGB1A peptide had a higher gene transfer efficiency than rHMGB1A. Finally, rTAT-HMGB1A had no cytotoxicity to HEK 293 cells suggesting that rTAT-HMGB1A may be useful as a non-toxic gene delivery carrier.
- Subjects
NUCLEIC acids; GENETIC transformation; DNA; CARRIER proteins; GENE expression; CHROMATOGRAPHIC analysis; BIOLOGICAL assay; PEPTIDES; CELL-mediated cytotoxicity
- Publication
Biotechnology Letters, 2008, Vol 30, Issue 8, p1331
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-008-9695-4