Stretching DNA Fibers out of a Chromosome in Solution Using Electro-osmotic Flow

This research develops a bio-nanotechnology for mechanically handling individual DNA fibers. The current study particularly focuses on surgery of a chromosome, which is a long strand of DNA tightly wound on proteins. Since DNA wound on a chromosome is several centimeter long, it is too long to observe under a microscope. Therefore, it is necessary to develop a method for partial unwinding. This work uses laser-induced local heating. Focusing a laser beam onto an aimed location on a chromosome immersed in protease solution, induces temperature to rise and locally activates the enzymatic reaction. This work demonstrates stretching DNA fibers out of a chromosome using electro-osmotic flow. Results show a chromosome immobilized onto a glass surface with released DNA fibers as long as 150μm.

[1]  M. Washizu,et al.  Electrostatic manipulation of DNA in microfabricated structures , 1989, Conference Record of the IEEE Industry Applications Society Annual Meeting,.

[2]  D. Ward,et al.  Immunological method for mapping genes on Drosophila polytene chromosomes. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Jan Greve,et al.  Unfolding individual nucleosomes by stretching single chromatin fibers with optical tweezers , 2001, Nature Structural Biology.

[4]  Masao Washizu,et al.  Stretching yeast chromosomes using electroosmotic flow , 2003 .

[5]  C. Bustamante,et al.  An integrated laser trap/flow control video microscope for the study of single biomolecules. , 2000, Biophysical journal.

[6]  V. Croquette,et al.  Replication by a single DNA polymerase of a stretched single-stranded DNA. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. J. Hunter.  Zeta potential in colloid science : principles and applications , 1981 .

[8]  H. Schreier,et al.  Overexpression and characterization of a prolyl endopeptidase from the hyperthermophilic archaeon Pyrococcus furiosus , 1997, Journal of bacteriology.

[9]  Pei-Hsing Huang,et al.  Molecular dynamics analyses of the femtosecond laser-induced grain boundary spallation , 2007 .

[10]  H. Heng,et al.  High-resolution mapping of mammalian genes by in situ hybridization to free chromatin. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Akira Mizuno,et al.  Manipulation of single coiled DNA molecules by laser clustering of microparticles , 2002 .

[12]  S. Smith,et al.  Single-molecule studies of DNA mechanics. , 2000, Current opinion in structural biology.