Rapid gene cloning using terminator primers and modular vectors.
We seek to create useful biological diversity by exploiting the modular nature of genetic information. In this report we describe experiments that focus on the modular nature of plasmid cloning vectors. Bacterial plasmids are modular entities composed of origins of replication, selectable markers and other components. We describe a new ligation-independent cloning method that allows for rapid and seamless assembly of vectors from component modules. We further demonstrate that gene cloning can be accomplished simultaneously with assembly of a modular vector. This approach provides considerable flexibility as it allows for 'menu driven' cloning of genes into custom assembled modular vectors.
Donahue
William F
WF
Department of Pharmacology and Experimental Therapeutics, Boston University Medical Center, 80 East Concord Street, Boston, MA 02118, USA.
Turczyk
Brian M
BM
Jarrell
Kevin A
KA
D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012265:Ribonucleotides / Q000235:genetics / Q000378:metabolism; D013997:Time Factors
D017931:DNA Primers; D005819:Genetic Markers; D012265:Ribonucleotides; C072146:Pfu DNA polymerase; D004259:DNA-Directed DNA Polymerase