| 摘要: |
| 据已发表的CSFV C-株序列，设计合成了5′-UTR特异性PCR引物对P1-NI及半套式下游引物N1′。从兔脾组织中提取总RNA，应用RT-PCR及Half-nested PCR成功地扩增出CSFV 5′-UTR，进一步将该片段克隆到pMD-18T载体质粒并进行了序列测定。与Shimen株、ALD株、GPE-株、Holland C-株和Russian C-株相应区段的比较分析表明，其同源性分别为96.2%,95.7%,96.0%,98.7%和95.4%。5′-UTR中发现的隐性AUG超始密码子及茎环二级结构，可能在多聚蛋白前体的翻译以及病毒的复制中起着重要作用。 |
| 关键词: 猪瘟 中国猪瘟兔化弱毒 5′-UTR 克隆 序列及二级结构分析 |
| DOI： |
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| 基金项目:国家重点基础研究发展规划项目(G19990119) |
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| Cloning and secondary structure analysis of genomic 5＇-UTR of Chinese lapnized attenuative strain(derived from spleen) |
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| Abstract: |
| Based on published sequences of strain-C of CSFV,a specific PCR prim er pair P1-N1 and a half-nested PCR reverse prim er N1′were designed.Utilizing RT-PCR and half-nested PCR techniques,genom ic 5′-UTR of strain-C was successfully amplified after the total RNA had been extracted from spleensof experimentally infected rabbits.The target fragmentwas further sequenced after being cloned into pMD 18-T vector plasmid.Sequence comparison with corresponding regions of other strain srevealed that the nucleotide sequence homologies of 5′-UTR of strain-Care 96.2% with Shimen-strain,95.7% with ALD-strain,96.0% with GPE-strain, 98.7% with Holland C-strain, and 95.4% with Russian C-strain.Several cryptic AUG start codons (CASCs) and stem-loop secondary structures in the 5′-UTR perhaps play an important role in the process of pre-polyprotein translation and viral replication. |
| Key words: classical swine fever virus Chinese lapnized attenuative strain of CSFV 5′-UTR cloning sequence and secondary structure analysis |
