摘要
[目的]探索检测苹果褪绿叶斑病毒的技术体系。[方法]利用CTAB法提取感病苹果叶片总RNA,以总RNA为模板,利用特异性引物,建立利用cDNA探针检测苹果褪绿叶斑病毒的技术体系。[结果]扩增出了预期片段,经克隆、测序证明该片段为苹果褪绿叶斑病毒的特异性片段。以带有特异片段的质粒为模板,利用PCR技术制备出地高辛标记的cDNA探针。[结论]以质粒为模板可高效制备大量探针。地高辛标记探针为非放射性标记探针,安全性较好,灵敏度较高,具有重要的应用价值。
[ Objective ] The aim was to develop a system of detection for Apple chorotic leaf spot virus （ACILSV） with molecular hybridiza- tion. [ Method ] Total RNA was extracted from the leaves of infected apple by CTAB method, and the total RNA was used as the template to amplify the genome of ACLSV with the ACLSV specific primers. The system of detection of ACI.SV was developed by eDNA probe. [ Result] The predicted fragment was obtained, and the specific fragment was verified by cloning and sequencing. The digoxigenin-labeled eDNA probe is prepared by PCR using the plasmid with the specific fragment as a template. [ Conclusion ] The plasmid DNA was used as the template, then a number of probe could be effectively prepared. In addition, the digoxigenin-labeled probe was a non-radioactive one, so it was safe and was also high sensitive. The probe has an important significance for application.
出处
《安徽农业科学》
CAS
2013年第31期12324-12325,共2页
Journal of Anhui Agricultural Sciences
关键词
苹果褪绿叶斑病毒
CDNA探针
检测
Apple chorotic leaf spot virus
cDNA probe
Blot hybridization
Detection
