蓝舌病相关病毒EUBANANGEE RNA2及RNA3的cDNA克隆及序列分析

cDNA CLONING AND SEQUENCING OF EUBANANGEE YIRUS RNA SEGMENT 2 AND 3

本文报道了蓝舌病相关病毒Eubanangee RNA 2与RNA 3 的cDNA克隆及序列分析。采用随机引物将纯化的病毒的dsRNA的第二,三片段逆转录成cDNA,用Sau3AI消化,克隆于BamHI酶切的M_(13)载体中,然后用Sanger双脱氧链终止法进行核酸的序列分析。结果表明:RNA3与BTV(蓝舌病)WJRNA3的同源性达70％以上,而RNA 2与BTV的RNA 2同源性较小。

Eubanangee is a member of the orbivirus genus. In this paper we report the cDNA cloning and sequencing of Eubanangee virus RNA2 and RNA3. Viral dsRNA was purified and random primer was used to synthesize cDNA. The cDNA was digested with Sau3AI, cloned into Bam-HI digested Ml3 vector, Nucleotide sequence analysis was performed by using Sanger dideoxy chain-termination method. The results showed that the homology of Eubanangee RNA3 and blue tongue virus RNA3 was more than 70%at nucleotide level. The homology of RNA2 of two virus was very low, which indicate that the RNA2 is functionally neutralizing antibody.