%0 Journal Article
%T BglBrick vectors and datasheets: A synthetic biology platform for gene expression
%A Taek Lee
%A Rachel A Krupa
%A Fuzhong Zhang
%A Meghdad Hajimorad
%A William J Holtz
%A Nilu Prasad
%A Sung Lee
%A Jay D Keasling
%J Journal of Biological Engineering
%D 2011
%I BioMed Central
%R 10.1186/1754-1611-5-12
%X Here, we report the development and characterization of a library of expression vectors compatible with the BglBrick standard (BBF RFC 21). We have designed and constructed 96 BglBrick-compatible plasmids with a combination of replication origins, antibiotic resistance genes, and inducible promoters. These plasmids were characterized over a range of inducer concentrations, in the presence of non-cognate inducer molecules, and with several growth media, and their characteristics were documented in a standard format datasheet. A three plasmid system was used to investigate the impact of multiple origins of replication on plasmid copy number.The standardized collection of vectors presented here allows the user to rapidly construct and test the expression of genes with various combinations of promoter strength, inducible expression system, copy number, and antibiotic resistance. The quantitative datasheets created for these vectors will increase the predictability of gene expression, especially when multiple plasmids and inducers are utilized.Metabolic engineering, the redirection of metabolic pathways using genetic manipulation, plays an important role in a wide range of biological research including drug production, bioremediation, and biofuel production [1-5]. Metabolic pathways that lead to important drugs or chemicals are often multi-step processes involving many enzymes. In addition, controlling and coordinating the activity of each enzyme to achieve the optimal production of the target product is extremely complicated [6-9]. To construct an entire metabolic pathway in a heterologous host, the genes encoding the pathway enzymes often have to be constructed on multiple plasmids. Furthermore, the expression of each enzyme needs to be tuned to balance it with that of the other enzymes in the pathway and to reduce the metabolic burden on the host cell [6,9-11]. Recently, several advanced cloning methods using homologous recombination, such as Sequence and Ligation-Ind
%U http://www.jbioleng.org/content/5/1/12