[使用生物素化寡脱氧核糖核苷酸测定马铃薯纺锤块茎类病毒和菊花矮化类病毒]

[Determination of potato spindle tumor viroid and chrysanthenum stund viroid using biotinylated olideoxyribonucleotides].

作者信息

Meldraĭs Ia, Lebedeva I V, Druka A Ia, Ivanovskaia M G, Shabarova Z A, Gurinovich T I, Birznieks K A, Keks V G, Zeĭdaka A A

出版信息

Mol Biol (Mosk). 1992 May-Jun;26(3):540-5.

PMID:1406609

Abstract

A 26 base long oligodeoxyribonucleotide complementary to a common RNA sequence of potato spindle tuber viroid (PSTV) and chrysantemum stunt viroid (CSV) was synthesized. The 3'-end biotinylated one was used for the detection of PSTV and CSV RNA immobilized on nitrocellulose filters by nucleic acid hybridization. Visualization of hybridization results was performed by two ways, either by streptavidin-alkaline phosphatase conjugate or streptavidine and biotinylated alkaline phosphatase. It was possible to detect 0.65 ng of purified CSV and PSTV RNA. The suggested system of viroid diseases detection can be used by agricultural and horticultural enterprises.

摘要

合成了一段26个碱基长的寡聚脱氧核糖核苷酸，它与马铃薯纺锤块茎类病毒（PSTV）和菊花矮化类病毒（CSV）的常见RNA序列互补。将3'-末端生物素化的该寡聚脱氧核糖核苷酸用于通过核酸杂交检测固定在硝酸纤维素滤膜上的PSTV和CSV RNA。杂交结果的可视化通过两种方式进行，即使用链霉亲和素-碱性磷酸酶偶联物或链霉亲和素和生物素化碱性磷酸酶。能够检测到0.65 ng纯化的CSV和PSTV RNA。所建议的类病毒疾病检测系统可供农业和园艺企业使用。