title: [Construction of randomly matrix attachment regions library from the green alga: Dunaliella salina].
abstract: Matrix attachment region (MAR) is DNA fragment that can bind to the nuclear matrix. In order to isolate the MAR fragment from the halotolerant green alga Dunaliella salina, we created a library of randomly obtained MAR from D. salina. Firstly the intact nuclei were released using 0.5% Triton X-100, then purification was carried out by discontinuous centrifugation using 30% and 70% Percoll gradients. Histones of nuclear matrices were removed using 25mmol/L lithium dioodosalicylate, the DNAs not closely associated with the matrices were removed using restriction enzymes. The remained matrices DNAs were digested by proteinase K, extracted with phenol/chloroform and precipitated with ethanol, and then cut with four kinds of restriction enzymes, the resulting DNAs were subsequently ligated to pUC18-vector and transferred to E. coli JM109 strains, DNA sequencing showed that the DNA fragments had the features of MAR DNA fragments.
authors.affiliation: Laboratory for Cell Biology, Zhengzhou University 450052.
authors.forename: Tian Yun
authors.initials: TY
authors.lastname: Wang
authors.forename: Wei Hong
authors.initials: WH
authors.lastname: Hou
authors.forename: Bao Mei
authors.initials: BM
authors.lastname: Yuan
authors.forename: Yu Rong
authors.initials: YR
authors.lastname: Chai
authors.forename: Gui Qin
authors.initials: GQ
authors.lastname: Hou
authors.forename: Jian Min
authors.initials: JM
authors.lastname: Wang
authors.forename: Le Xun
authors.initials: le X
authors.lastname: Xue
mesh_terms: D000460:Chlorophyta / Q000235:genetics*; D048028:DNA, Algal / Q000235:genetics* / Q000302:isolation & purification*; D045170:Matrix Attachment Regions / Q000235:genetics*
chemical_list: D048028:DNA, Algal