Rapid DNA library construction for functional genomic and metagenomic screening.
A rapid protocol was developed for constructing plasmid libraries from small quantities of genomic/metagenomic DNA. The technique utilizes linker amplification with topoisomerase cloning and allows for inducible transcription in Escherichia coli. As proof of principle, several anti-Bacillus lysins were cloned from bacteriophage genomes and an aerolysin was cloned from a metagenomic sample.
Schmitz
Jonathan E
JE
Laboratory of Bacterial Pathogenesis and Immunology, Rockefeller University, 1230 York Avenue, Box 172, New York, NY 10021, USA.
Daniel
Anu
A
Collin
Mattias
M
Schuch
Raymond
R
Fischetti
Vincent A
VA
D001427:Bacterial Toxins / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli; D015723:Gene Library; D005820:Genetic Testing / Q000379:methods*; D023281:Genomics / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques; D010957:Plasmids / Q000235:genetics*; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics; D014158:Transcription, Genetic
D001427:Bacterial Toxins; D052899:Pore Forming Cytotoxic Proteins; C032676:aerolysin