Genomic modification of eukaryotes using BAC recombineering

Abstract

Introduction of modified genes into eukaryotic genome gives new opportunities for investigations of biological processes. Gene expression is controlled by specific regulator regions of different size and localization in genome. Most of them are poorly studied. Thus, a transgene construct is usually introduced together with a short ubiquity promoter region. Since all original regulator regions are lost, it makes difficulties to get a high level of expression and particularly a tissue-specific expression of transgene. This article describes a new method of genome modification using bacterial artificial chromosome (BAC). Method is based on recombination system of Rаc-prophage (Rec/ET) or λ-phage (Redα/Redβ) transferred into Escherichia coli DH10B cells. In case of BAC recombination, all known regulatory regions essential for desired transgene expression can be included. This method is used for generation of transgenic animals that allows more physiological or tissue-specific expression of transgene in vivo.

Keywords

• cloning
• bac recombineering
• gene expression
• genomic modification