Expression, Purification and DNA binding activity of recombinant human XRCC1: A DNA repair gene
Shagufta Zafar and Krishna Prakash*
Recombinant DNA Technology Laboratory, Department of Biotechnology, Central University of South Bihar, Panchanpur Road, Gaya - 824236 (India) Corresponding Author: Krishna Prakash* Email ID: krishna@cub.ac.in
ABSTRACT
The XRCC1 (X-ray repair cross-complementing protein 1) is a DNA repair protein involved in repairing of damaged DNA. It primarily repairs DNA damage caused by alkylation, ionizing radiation, and the genotoxic agents. It acts as a molecular scaffold protein associated with single-strand DNA break repair and base excision repair pathway proteins like, poly nucleotide kinase, DNA polymerase, poly (ADP- ribose) polymerase I, and DNA ligase III, etc. The XRCC1 Open Reading Frame (ORF) encodes a 633-amino-acid protein with a 69.6 kDa molecular mass. Recently, a central DNA Binding domain is identified in the XRCC1 protein, whose complete characterization is not completely done. Therefore, the aim of this study is to characterize the XRCC1 protein Central DNA Binding Domain by studying its DNA binding activity by the DNA agarose gel based EMSA (Electrophoretic Mobility Shift Assay) technique, which is essential for interaction at damage repair sites