论文部分内容阅读
目的设计版纳病毒型别特异性测序引物,为构建快速高效经济的版纳病毒全基因组测序平台奠定基础。方法根据Gen Bank已知的版纳病毒基因序列信息进行基因型分析,采用Primer Premier v6.0及Oligo 7.56软件完成版纳病毒亚型特异性扩增引物的设计与评估。使用中国分离的30株版纳病毒进行版纳病毒型别特异性引物工作效率验证和评估。结果通过对30株隶属于2个不同基因亚型的版纳病毒开展全基因组序列测定,获得2套型别工作效率高、扩增效果稳定的基因A型特异性版纳病毒全基因组序列扩增测序引物。基因A1亚型引物一套共26对;基因A2亚型引物一套共计30对。完成了30株版纳病毒全基因组序列扩增。结论设计的版纳病毒型别特异性扩增测序引物可以高效准确地完成版纳病毒全基因组序列扩增,为进一步深入研究版纳病毒分子生物学特征奠定了基础。
OBJECTIVE: To design a type-specific DNA sequencing primer for Banna virus and lay a foundation for constructing a fast and cost-effective platform for genome-wide sequencing of Banana virus. Methods Based on the GenBank sequence information of GenBank virus known by Gen Bank, Primer Premier v6.0 and Oligo 7.56 software were used to complete the design and evaluation of the specific amplification primers of the subgenotypes of the virus. 30 Banna viruses isolated in China were used to validate and evaluate the efficiency of Banna virus type-specific primers. Results Genomic sequence analysis of 30 GenBank viruses belonging to two different subtypes of genotypes revealed that genotype A specific full-length genomic sequence-amplified primers were obtained with high efficiency and stable amplification. . A total of 26 pairs of gene A1 subtype primers; a total of 30 pairs of gene A2 subtype primers. The complete genomic sequence amplification of 30 strains of Banna virus was completed. Conclusion The designed type-specific PCR primers can amplify the complete genome sequence of the BAV virus efficiently and accurately, which lays the foundation for further study on the molecular biological characteristics of the BAV virus.