title: Rapid screening for bacterial colonies harbouring tandem hepatitis B virus sequences by an oligonucleotide probe.
abstract: Transfection of the hepatitis B virus (HBV) genome requires the cloning of tandem HBV sequences into a plasmid vector, which is usually screened for by restriction enzyme digestion of plasmid minipreparations from at least a dozen bacterial colonies. We describe a simple alternative screening method based on in situ hybridization of bacterial colonies with a [32P]-labelled synthetic oligonucleotide which spans the head-to-tail junction site of two tandem HBV molecules. The accurate detection by the oligoprobe is confirmed by enzymatic digestion.
authors.affiliation: Unité de Recherche Sur Les Hépatites INSERM 271, Lyon, France.
authors.forename: S P
authors.initials: SP
authors.lastname: Tong
authors.forename: J S
authors.initials: JS
authors.lastname: Li
authors.forename: L
authors.initials: L
authors.lastname: Vitvitski
authors.forename: S
authors.initials: S
authors.lastname: Benjelloun
authors.forename: C
authors.initials: C
authors.lastname: Trépo
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D014162:Transfection / Q000235:genetics
chemical_list: D015345:Oligonucleotide Probes