Binding of low mobility group proteins with DNA examined in homologous and heterologous systems by Gel retardation assay

• Published in: Cell biology international Vol. 17; no. 1; pp. 45 - 53
• Main Authors: Wiland, Eva, Siemieniako, Barbara, Trzeciak, Wieslaw H.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier Ltd 1993; Blackwell Publishing Ltd; Elsevier
• Subjects: Animals; Antigens, Neoplasm - genetics; Base Sequence; Biological and medical sciences; Chromatin - chemistry; DNA, Neoplasm - analysis; DNA, Neoplasm - genetics; DNA, Neoplasm - metabolism; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; interaction with DNA; LMG proteins; LMG proteins, interaction with DNA; Miscellaneous; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Molecular Weight; Neoplasm Proteins - analysis; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Oligonucleotides; Pancreatic Neoplasms - chemistry; Pancreatic Neoplasms - immunology; Pancreatic Neoplasms - pathology; Protein Binding; Rats; Tumor Cells, Cultured; TBE, Tris:borate:borate:EDTA; SDS, sodium dodecylsulphate; PMSF, phenyl methylsulfonylfluoride; LMG proteins, interaction with DNA; PBS, phosphate buffered saline; TCA, trichloroacetic acid; LMG, low mobility group; BSA, Bovine serum albumin; HMG, high mobility group; HEPES (N-[2-hydroxyethyl] piperazine-N′-[2-ethanesulfonic acid]); DTT, dithiothreitol; Heterologous system; Carcinoma; Gel electrophoresis; Interaction; Homologous system; Gel retardation technique; Malignant tumor; Fragment; Fixation; Cell line; Gene; DNA; Pancreas
• ISSN: 1065-6995; 1095-8355
• DOI: 10.1006/cbir.1993.1004

The interaction of low mobility group proteins (LMG), isolated from chromatin of pancreatic carcinoma cells (CAPAN-2), with fragments of 5′-flanking region of the antigen 17-1A gene was studied by gel retardation assay. The LMG proteins, which formed complexes with DNA were extracted from the gels and identified by polyacrylamide gel electrophoresis under denaturing conditions. The proteins of Mw about 100, 60, 55 and 48 kDa, which formed specific complexes with fragments of 5′-flanking region of the antigen 17-1A gene, were identified.