Rapid screening for bacterial colonies harbouring tandem hepatitis B virus sequences by an oligonucleotide probe.
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.
Tong
S P
SP
Unité de Recherche Sur Les Hépatites INSERM 271, Lyon, France.
Li
J S
JS
Vitvitski
L
L
Benjelloun
S
S
Trépo
C
C
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
D015345:Oligonucleotide Probes