Open AccessEXPRESSION AND PURIFICATION OF RECOMBINANT T4 DNA LIGASE IN E. COLI
Author(s) -
Duong Long Duy,
Duc Van Luong,
Hieu Thi Phuong Nguyen,
Hương Thanh Trần,
Thao Thi Phuong Dang,
Thuoc Linh Tran
Publication year - 2011
Publication title -
khoa học công nghệ
Language(s) - English
Resource type - Journals
ISSN - 1859-0128
DOI - 10.32508/stdj.v14i3.1991
Subject(s) - dna ligase , recombinant dna , microbiology and biotechnology , lac operon , plasmid , dna , biology , western blot , escherichia coli , dna ligases , myc tag , affinity chromatography , chemistry , gene , biochemistry , enzyme , fusion protein
In this study, we report results on the expression and purification of recombinant T4 DNA ligase. Plasmid pET16b-T4Dnl contains the gp30 gene which encodes for T4 DNA ligase. The target protein is fused with 10xHis tag to facilitate the purification and recovery. pET16b-T4Dnl was transformed into E. coli BL21(DE3) and then induced the expression of 10xHis-T4Dnl by IPTG. The recombinant protein was purified by Ni-NTA chromatography and confirmed by SDS-PAGE and Western blot. The activity of purified protein was tested by joining DNA λ/HindIII.