New bacteriophage vectors for the large scale production of single stranded insert DNA.
SSEV 18 and SSEV 19, derivatives of the bacteriophages M13mp18/19, are new versatile cloning vectors allowing the large scale preparation of single stranded (ss) insert DNA. Replacing the original multiple cloning site by a synthetic 96 bp DNA fragment, a new polylinker region has been introduced containing complementary sequences designed to form a stem structure where single stranded insert fragments can be excised via a 'master restriction' site. The usefulness of such a vector has been demonstrated by the cloning of a 900 bp HindIII fragment derived from the Mycoplasma hyorhinis 23 S rRNA gene. After excision the single stranded insert was labeled isotopically and tested for sensitivity and specificity in detecting homologous sequences in pure DNA or cellular material immobilized on filters.
Biernat
J
J
Biosyntech Biochemische Synthesetechnik GmbH, Hamburg, F.R.G.
Göbel
U B
UB
Köster
H
H
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D003090:Coliphages / Q000235:genetics*; D004251:DNA Transposable Elements; D004275:DNA, Ribosomal / Q000235:genetics; D004277:DNA, Single-Stranded / Q000138:chemical synthesis / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D012335:RNA, Ribosomal / Q000235:genetics; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D004275:DNA, Ribosomal; D004277:DNA, Single-Stranded; D009838:Oligodeoxyribonucleotides; D012335:RNA, Ribosomal