Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Bacterial Toxins / genetics
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Cloning, Molecular
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Escherichia coli
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Gene Library*
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Genetic Testing / methods*
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Genomics / methods*
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Nucleic Acid Amplification Techniques
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Plasmids / genetics*
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Pore Forming Cytotoxic Proteins / genetics
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Transcription, Genetic
Substances
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Bacterial Toxins
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Pore Forming Cytotoxic Proteins
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aerolysin