Cloning And Characterization Of Full-length CDNA Fragment Of Gene Encoding MAR-binding Protein From Dunaliella Salina

Matrix attachment regions (MARs) are important boundary elements in eukaryotic genome, which still bind to the nuclear matrices after chromatin digested with restriction endonuclease. It has been demonstrated that MARs increase transgene expression. By means of interacting with some MARs, MAR-binding proteins play a key role in eukaryotic gene replication, expression, regulation and chromosomal construction. So far, some MAR-binding proteins have been isolated from yeast, plants, animals and human, but not from unicellular eukaryotic algae.Dunaliella salina (D. salina), an autotropic unicellular eukaryotic green alga, which belongs to Chlorophyta, Chlorophyceae, Volvocales. It is one of the most extremely halotolerant eukaryotes and can live in a variety of salt concentrations ranging from 0.05mol/L to 5mol/L solution of sodium chloride. The aim of this study is to clone MAR-binding proteins from D. salina, provide some experimental bases for the study of the effect and mechanism of MAR-binding proteins, and do some exploring for the establishing the steady and effective D. salina expression systems.Here, A pair of degenerate primers was designed according to the homologous amino acid sequences conserved motifs of other organisms known MAR-binding protein, and then used to amplify MAR-binding protein cDNA fragment from D. salina by PCR. According to the known cDNA sequence of MAR-binding protein from D. salina, 5'RACE and 3'RACE were performed to obtain 5' and 3' ends sequences of...