Lebedeva I V, Ivanovskaia M G, Gurinovich T I, Gottykh M B, Meldraĭs Ia A, Shabarova Z A
Bioorg Khim. 1993 Sep;19(9):894-904.
A non-radioactive diagnosticum for plant viroid diseases has been designed, based on hybridization of a biotin-labeled 26-member oligonucleotide probe with the viroid RNA site identical for potato spindle tuber viroid and chrysanthemum stunt viroid. The biotin label has been introduced into the synthetic oligonucleotide probe by the high-yield acylation of the oligonucleotide aminoalkylamide with the biotin imidazolide or N-hydroxysuccinimide ester. Hybridization techniques have been elaborated for nucleic acids isolated from plant sap. The hybrids obtained have been detected with streptavidin and biotinylated alkaline phosphatase or with the covalent conjugate of streptavidin and alkaline phosphatase, the sensitivity being as low as 1 ng. The methodology used can be applied for revealing viroids and for large scale and quick investigation of plant cell cultures.
基于生物素标记的26个成员的寡核苷酸探针与马铃薯纺锤块茎类病毒和菊花矮缩类病毒相同的类病毒RNA位点杂交，设计了一种用于植物类病毒疾病的非放射性诊断试剂。通过寡核苷酸氨基烷基酰胺与生物素咪唑化物或N-羟基琥珀酰亚胺酯的高产率酰化反应，将生物素标记引入合成寡核苷酸探针中。已经为从植物汁液中分离的核酸精心设计了杂交技术。所获得的杂交体已用抗生物素蛋白和生物素化碱性磷酸酶或抗生物素蛋白与碱性磷酸酶的共价缀合物进行检测，灵敏度低至1纳克。所使用的方法可用于检测类病毒以及大规模快速研究植物细胞培养物。
