Bacterial Artificial Chromosomes

Abstract

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Overcoming the Challenges of Megabase-Sized Plasmid Construction in Escherichia coli

“…Plasmid and artificial chromosome vectors have been used to carry genes, gene clusters, genomic regions, and even a whole bacterial genome. − There is a huge demand for a useful cell-based platform for megabase-sized plasmid construction in the era of genome writing projects . Traditionally, the yeast artificial chromosome (YAC) vector has been used for assembling megabase-sized synthetic chromosomes. − A hybrid of the bacterial artificial chromosome (BAC) vector and YAC, or BAC-YAC, enabled the shuttling of up to 0.5 Mb chromosomes from yeast to Escherichia coli . Larger synthetic bacterial chromosomes built in yeast are yet to be transplanted into bacterial cells. , Another approach is to directly use E. coli and BAC for constructing sub-megabase-sized chromosomes. , The E. coli platform would hopefully become a good alternative to yeast, because E. coli is able to transfer anywhere from over 100 kbp up to a few megabase-sized DNA to many kinds of cells via conjugation machinery. − …”

“…It has been known for decades that the traditional BAC vector derived from the E. coli F plasmid has ability to carry moderately large DNA fragments (up to 350 Kb). In the most successful case, E. coli was transformed with 457 Kb and 513 Kb BAC-YACs .…”