title: Characterisation of synthetic DNA probe detecting potato spindle tuber viroid.
abstract: Chemically synthesized DNA fragments complementary to selected regions of the potato spindle tuber viroid (PSTV) genome were cloned into Escherichia coli plasmid pUC9. One of the recombinant plasmids (pIBB4) with a 87 bp insert representing the central region of the PSTV genome (nucleotides 88 to 174) was used after labelling by nick translation for detecting PSTV by dot-blot hybridization. The molecular probe was almost as sensitive as the one carrying the full genomic PSTV copy (pAV401), detecting down to 20 pg of PSTV RNA in 8-15 micrograms of infected tissue. The specificity of the test was high; no signals were created by extracts from healthy plants or plants infected with a variety of common potato viruses. The probe is potentially useful in studies on mixed infections of potatoes with different viruses, and in selection of certified seed material.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw.
authors.forename: M
authors.initials: M
authors.lastname: Wełnicki
authors.forename: J
authors.initials: J
authors.lastname: Skrzeczkowski
authors.forename: A
authors.initials: A
authors.lastname: Sołtyńska
authors.forename: P
authors.initials: P
authors.lastname: Jończyk
authors.forename: W
authors.initials: W
authors.lastname: Markiewicz
authors.forename: R
authors.initials: R
authors.lastname: Kierzek
authors.forename: B
authors.initials: B
authors.lastname: Imiołczyk
authors.forename: W
authors.initials: W
authors.lastname: Zagórski
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000138:chemical synthesis / Q000235:genetics*; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000302:isolation & purification; D010942:Plant Viruses / Q000235:genetics / Q000302:isolation & purification*; D010957:Plasmids; D012367:RNA, Viral / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000382:microbiology*; D013045:Species Specificity; D014772:Viroids / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D012367:RNA, Viral; D004247:DNA