中国猪瘟兔化弱毒全基因组cDNA文库的构建和序列分析

Construction of genomic cDNA library and sequence analysis of the lapinized attenuated CSFV C strain

根据已发表的猪瘟病毒 (CSFV)序列设计并合成了 2 6条覆盖全长基因组的套式和半套式PCR引物。应用RT PCR、NestedPCR和Half nestedPCR技术从试验感染的兔脾中成功地扩增出了各相应的cDNA重叠片段 ,分别克隆和测序 ,构建了C株全基因组cDNA文库。采用DNAstar软件对全基因组的核苷酸和氨基酸序列进行了同源性比较分析。CS

Based on the published sequences of CSFV , 26 nested and half nested PCR primers covering the genome sequence of C strain were designed. Utilizing the RT PCR, nested and half nested PCR techniques, corresponding overlaping cDNA fragments were successfully amplified from total RNA extracted from spleens of experimentally infected rabbits. C strain of CSFV genomic cDNA library was constructed after cloning and sequencing of thesefragments. Comparing and analysing of homologies of nucleotideand amino acid sequences were performed by computer software DNAstar. Construction of genomic cDNA library makes the basis for construction of infectious full length cDNA of CSFV C strain.