TAILIEUCHUNG - Báo cáo khoa hoc : Genome walking in eukaryotes

Genome walking is a molecular procedure for the direct identification of nucleotide sequences from purified genomes. The only requirement is the availability of a known nucleotide sequence from which to start. | fFEBS Journal REVIEW ARTICLE Genome walking in eukaryotes Claudia Leoni1 Mariateresa Volpicella1 Francesca De Leo2 Raffaele Gallerani1 and Luigi R. Ceci2 1 Department of Biochemistry and Molecular Biology University of Bari Italy 2 Institute of Biomembranes and Bioenergetics Italian NationalResearch Council Bari Italy Keywords gene tagging gene therapy gene trapping genome walking insertional mutagenesis multigene family next generation sequencing T-DNA transposable elements virus integration Correspondence L. R. Ceci Institute of Biomembranes and Bioenergetics CNR Via Amendola 165 A 70126 Bari Italy Fax 39 080 544 3317 Tel 39 080 544 3311 E-mail Received 1 April 2011 accepted 12 August 2011 doi Genome walking is a molecular procedure for the direct identification of nucleotide sequences from purified genomes. The only requirement is the availability of a known nucleotide sequence from which to start. Several genome walking methods have been developed in the last 20 years with continuous improvements added to the first basic strategies including the recent coupling with next generation sequencing technologies. This review focuses on the use of genome walking strategies in several aspects of the study of eukaryotic genomes. In a first part the analysis of the numerous strategies available is reported. The technical aspects involved in genome walking are particularly intriguing also because they represent the synthesis of the talent the fantasy and the intelligence of several scientists. Applications in which genome walking can be employed are systematically examined in the second part of the review showing the large potentiality of this technique including not only the simple identification of nucleotide sequences but also the analysis of large collections of mutants obtained from the insertion of DNA of viral origin transposons and transfer DNA T-DNA constructs. The enormous amount of data obtained indicates that

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