YAC and BAC cloning systems

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Presentation transcript:

YAC and BAC cloning systems YAC and BAC cloning systems. The general steps involved in the construction of yeast artificial chromosome (YAC, left) and bacterial artificial chromosome (BAC, right) clones are summarized. Specifically, high-molecular-weight source DNA (e.g., human DNA) is carefully prepared, partially digested with a restriction enzyme, and size selected to yield large DNA fragments (e.g., typically about 200 to 1000 kb for YACs and about 100 to 300 kb for BACs). Appropriate vector sequences are then ligated to the size-selected, insert DNA. For YACs, this consists of two vector arms that together contain all the structural elements necessary for the propagation of a chromosome in yeast (see Green et al.76 for details). For BACs, this consists of a single vector fragment that contains a suitable antibiotic-resistance gene (see Birren et al.114 for details). The ligated DNA is then transformed into appropriately prepared yeast or bacterial cells, respectively. The systems are set up such that the only cells that grow are those containing the appropriate yeast-selectable markers (in the case of YACs) or antibiotic-resistance gene (in the case of BACs). Note that the resulting YACs and BACs are linear and circular DNA molecules, respectively. Source: General Themes, The Online Metabolic and Molecular Bases of Inherited Disease Citation: Valle D, Beaudet AL, Vogelstein B, Kinzler KW, Antonarakis SE, Ballabio A, Gibson K, Mitchell G. The Online Metabolic and Molecular Bases of Inherited Disease; 2014 Available at: http://ommbid.mhmedical.com/DownloadImage.aspx?image=/data/Books/971/ch10fg2.png&sec=62633187&BookID=971&ChapterSecID=62632383&imagename= Accessed: October 20, 2017 Copyright © 2017 McGraw-Hill Education. All rights reserved