摘要
采用黄瓜花叶病毒(CMV)亚组Ⅰ株系Fny-CMVRNA_2的1209~1626核苷酸片段和亚组Ⅱ株系Ls-CMVRNA_2的2002~2433核苷酸片段的cDNA克隆,体外转录,同时掺入 ̄(32)P获得负链RNA探针,与纯化的番茄和甜椒上的CMV中国分离物的RNA杂交,结果表明:CMV番茄和甜椒中国分离物与Fny-CMV的核苷酸有高度同源性,隶属于Fny-CMV为代表的亚组Ⅰ株系。并利用K-CMV株系(亚组Ⅰ,源于中国)的RNA_2全长cDNA克隆的两个EcoRI位点间的核苷酸序列(1657~2125nt)作探针,与上述两种CMV中国分离物的RNA杂交,进一步比较分析了这两个分离物和K-CMV株系的关系。讨论了核酸酶保护法在CMV株系鉴定中的作用。
he cDNA sequences spanning nt 1209 to nt 1626 of Fny-CMV RNA2(a subgroupⅠstrain)and nt 2002 to nt 2433 of Ls-CMV RNA2(a subgroup Ⅱ strain)were used for probe preparation.The 32P-labeled negative strand RNAs prepared by transcription in vitro were hybridized with pu-rified RNAs of two Chinese CMV isolates from pepper and tomato,respectively.The resultshows that the two Chineses isolates share high identities of sequence to the Fny-CMV,thereforethe isolates belong to the CMV subgroupⅠ strain.A transcription probe complementary to thefragment flanking nt 1657 to nt 2125 of K-CMV RNA2(a subgroupⅠstrain originated from Chi-na) was hybridized with the purified RNAs of the two Chinese CMV isolates as well.The rela-tionship between the K-CMV strain and the two Chinese isolates was compared. The applicationof ribonuclease protection assay was discussed in the identification of CMV strains.
出处
《中国病毒学》
CSCD
1996年第1期69-76,共8页
Virologica Sinica