摘要
BVDV(牛病毒性腹泻病毒)是一类有重要经济价值的病毒,其核酸为单链RNA。该病毒提纯以后.用苯酚-氯仿-异戊醇提取RNA.在提取的BVDy RNA中所含的DNA杂质用DNase I降解除去,然后进一步用oligo dT纤维素拄亲和层析,琼脂糖凝胶电泳等方法纯化。纯化的BVDV RNA在E.coli,poly A聚合酶的催化作用下,在3’羟基末端聚腺化。cDNA在逆向转录酶的作用下,用polyA接尾的BVDV RNA作模板,oligodT_12—18作引物合成。琼脂糖凝胶电泳结果说明它与BVDV RNA相似。该cDNA的探针用斑点杂交方法与BVDV RNA,HCV RNA和酵母tRNA杂交,BVDV RNA和HCVRNA显阳性反应,酵母tRNA显阴性反应,结果说明合成的cDNA为BVDV RNA的cDNA。BVDV和HCV同为披盖科疫病毒属成员,它们具有部分相同的抗原,因此编码病毒蛋白的RNA序列具有同源序列,本研究证实了这种假设。
The BVDV(Bovine viral diarrhea virus)RNA is ssRNA.After the virus was purified, the BVDV RNA was extracted hy phenol-chloroform-isoamyl aleohl. The DNA contamination in the extracted BVDV RNA was degraded by DNase l. BVDV RNA was further purified by oligo dTcellulose affinity chromotography and agarose gel eleetrophoresis The purified BVDV RNA was polyadenylated at the 3′end under the catalysis of poly A polymerase, eDNA was synthesized with reverse transcriptase using the poly A-tailed BVDV RNA as template and oligo dT_(12-18) as primer The result of agarose gel electrophoresis of eDNA showed that it was similar to its template of BVDV RNA.The cDNA probe hybridized with BVDV RNA, HCV RNA and yeast tRNA by the dot blot method The BVDV RNA and HCV RNA were positive and yeast tRNA was negative. These results showed that synthetic eDNA was the eDNA of BVDV RNA BVDV and HCV are members of the pestivirus in togaviridae They had some identical antigens, so the sequences of RNA coded for viral proteins had the homologous parts of nucleotide sequences The studies proved this conjecture.
出处
《中国病毒学》
CSCD
1991年第3期242-247,共6页
Virologica Sinica
关键词
牛腹泻病毒
互补DNA
分子杂交
Bovine viral diarrhea virus Hog cholora virus Complementary DNA
