摘要
利用根据ToRSV加拿大悬钩子株系核酸序列设计、合成的寡核苷酸引物进行PCR扩增试验,能成功地扩增ToRSV悬钩子株系的目标片段,但不能扩增苹果、樱桃株系的目标片段。 而ToRSV苹果株系的克隆pS_(64),经EcoRI酶切,再分别用^(32)P—UTP、Dig—11—UTP和SP_6RNA多聚酶转录出^(32)P标记、Dig标记的cRNA探针后,不仅能有效地检测出ToRSV的苹果株系,而且能有效地检测樱桃株系和悬钩子株系。^(32)P与Dig—标记的探针灵敏度基本相同,而Dig标记的探针无放射性,可以长期保存,多次使用,灵敏、准确、安全、经济,是值得推广应用的检疫新方法。
Oligonucleotide primers corresponding to the sequence of Canadian raspberry strain were synthesized. With the polymerase chain reaction (PCR) , these primers successfully amplified the target fragment in ToRSV raspberry strain, but not in apple strain and cherry strain.Clone pS64 containing ToRSV apple strain RNA was transcribed to P32-labelled cRNA probe and dig -labelled cRNA probe separately. Both probes could detect apple, cherry and raspberry strains of ToRSV effectively and were equally sensitive. Dig - labelled probe is nonradioactive, long -stored, reuseable, sensitive, accurate, safe and economical. It is a new quarantine technique that can be extended and applied.
