Zhikhareva I V, Medvedeva T E, Aleksandrova G I, Klimov A I
Genetika. 1993 May;29(5):853-61.
The phenotype and localization of ts mutations in genomes of the influenza A/Victoria/30-ir (A/Vic/30-ir) and A/Hong Kong/17-ir (A/HK/17-ir) cold-adapted (ca) viruses were studied. Using the recombination analysis in chick embryo fibroblasts (CEF) we determined that influenza A/HK/17-ir ca virus carries ts mutations in three "internal" genes, i.e., PB1, NP and M, and influenza A/Vic/30-ir ca virus carries ones in four genes, i.e., PA, NP, M and NS. We have revealed ts mutations for NA gene in none of these viruses. Prior to the analysis of ts mutations in HA gene of influenza A/HK/17-ir and A/Vic/30-ir ca viruses, three cloning steps were performed in chick embryos (CE) by the method of limiting dilutions at 34 degrees C followed by selection of some strains with the most prominent ts phenotype. The cloned strains with such phenotypes were shown to repeat stable results within the recombination analysis in CE, i.e., none from the cloned strains of A/HK/17-ir ca virus recombined in CE at 40 degrees C with the 46 ts mutant, while recombination of this mutant with the cloned A/Vic/30-ir ca strains led to formation of the ts progeny. Thereafter our data result in conclusion that ts mutations in the PA gene must lead to some insignificant contribution for the expression of general ts phenotype among the ca strains as far as this sign is clearly displayed by both viruses, although only one of them, i.e., A/HK/17-ir carries ts mutation in the HA gene.
对甲型流感病毒A/维多利亚/30-ir(A/Vic/30-ir)和A/香港/17-ir(A/HK/17-ir)冷适应(ca)病毒基因组中的温度敏感(ts)突变的表型和定位进行了研究。通过在鸡胚成纤维细胞(CEF)中进行重组分析,我们确定甲型流感病毒A/HK/17-ir ca病毒在三个“内部”基因即PB1、NP和M中携带ts突变,而甲型流感病毒A/Vic/30-ir ca病毒在四个基因即PA、NP、M和NS中携带ts突变。我们在这些病毒中均未发现NA基因的ts突变。在分析甲型流感病毒A/HK/17-ir和A/Vic/30-ir ca病毒HA基因中的ts突变之前,通过在34℃下有限稀释法在鸡胚(CE)中进行了三个克隆步骤,随后选择了一些具有最显著ts表型的毒株。结果表明,具有这种表型的克隆毒株在CE中的重组分析中能重复稳定的结果,即甲型流感病毒A/HK/17-ir ca病毒的克隆毒株在40℃下于CE中均未与46 ts突变株发生重组,而该突变株与克隆的甲型流感病毒A/Vic/30-ir ca毒株的重组则导致ts子代的形成。此后,我们的数据得出结论,PA基因中的ts突变对ca毒株总体ts表型的表达必定只产生一些微不足道的影响,因为两种病毒均清晰显示出这一特征,尽管其中只有一种病毒即甲型流感病毒A/HK/17-ir在HA基因中携带ts突变。