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使用生物素标记的cDNA探针检测传染性法氏囊病病毒。

The use of biotin-labeled cDNA probes for the detection of infectious bursal disease viruses.

作者信息

Jackwood D J, Kibenge F S, Mercado C C

机构信息

Food Animal Health Research Program, Ohio Agricultural Research and Development Center, Ohio State University, Wooster 44691.

出版信息

Avian Dis. 1990 Jan-Mar;34(1):129-36.

PMID:2157388
Abstract

A cDNA library was prepared from the double-stranded RNA genome of the infectious bursal disease virus (IBDV) strain ST-C. The cDNA molecules were annealed into the plasmid pUC9 and used to transform Escherichia coli strain JM107. A cDNA clone that contained IBDV-specific nucleotide sequences was selected and designated STC-1. Radiolabeled probes were prepared from STC-1 and hybridized to genome segment A of ST-C in a northern blot hybridization assay. The STC-1 cDNA was 448 base pairs in length, and its nucleotide sequence indicated that it is located near the VP-2/VP-4 junction in IBDV genome segment A. Biotin-labeled probes were prepared from STC-1 and used in a dot-blot hybridization assay to detect IBDV. Under relatively low stringency conditions of hybridization, the biotinylated probes detected four subtypes of IBDV serotype 1 and a serotype 2 IBDV isolate.

摘要

从传染性法氏囊病病毒(IBDV)ST-C株的双链RNA基因组制备了一个cDNA文库。将cDNA分子退火到质粒pUC9中,并用于转化大肠杆菌JM107株。选择了一个含有IBDV特异性核苷酸序列的cDNA克隆,并将其命名为STC-1。从STC-1制备放射性标记探针,并在Northern印迹杂交试验中与ST-C的基因组A片段杂交。STC-1 cDNA长度为448个碱基对,其核苷酸序列表明它位于IBDV基因组A片段的VP-2/VP-4连接处附近。从STC-1制备生物素标记探针,并用于斑点印迹杂交试验以检测IBDV。在相对低严格度的杂交条件下,生物素化探针检测到IBDV血清型1的四个亚型和一个血清型2的IBDV分离株。

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