[腺病毒7型疫苗株左端0-17.5μ的克隆及0-4.8μ的测序]

[Cloning of 0-17.5 mu and sequencing of 0-4.8 mu of the lefe and DNA of the Ad7 vaccine strain left end].

作者信息

Shi C, Liu Y, Wen L

机构信息

Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 1999 Jun 30;13(2):136-8.

PMID:12569780

Abstract

OBJECTIVE

Cloning of 0-17.5 mu DNA fragment of adenovirus 7 vaccine strain and sequencing of 0-4.8 mu fragment (1,737 bp).

METHODS

Isolating and purifying Ad7 vaccine genome from the A549 cultured cells, putting 0.3-17.5 mu fragment into pAd7T plasmid, then sequencing the 0-4.8 mu fragment including inverted terminal repeats (ITR), packaging sequence and Ela region.

RESULTS

We obtained 0-17.5 mu fragment of Ad7 vaccine strain genome and sequenced its left terminal 1,737 bp. Sequence analysis showed that the Ad7 vaccine strain Ela region encodes 6,300, 24,000 and 28,000 proteins. Compared with equivalent region of Ad7 Gomen strain, they share the homology of the nucleotide sequence 98.9%, 97.3%, 97.5% and the homology of the deduced amino acid sequence 96.6%, 96.5%, and 96.9% respectively. When compared with Ad7 Grider strain, they share the homology of the nucleotide sequence 100%, 99.7%, 99.7% and the homology of the deduced amino acid sequence 100%, 99.1%, and 99.2% respectively.

CONCLUSION

The Ad7 vaccine strain left terminal 1,737 bp nucleotide sequence showed a high homology with corresponding region of Ad7 Gomen strain and Grider strain.

摘要

目的

克隆腺病毒7型疫苗株0 - 17.5 μ片段并对0 - 4.8 μ片段（1737 bp）进行测序。

方法

从A549培养细胞中分离纯化Ad7疫苗基因组，将0.3 - 17.5 μ片段插入pAd7T质粒，然后对包括反向末端重复序列（ITR）、包装序列和E1a区域的0 - 4.8 μ片段进行测序。

结果

获得了Ad7疫苗株基因组的0 - 17.5 μ片段，并对其左末端1737 bp进行了测序。序列分析表明，Ad7疫苗株E1a区域编码6300、2,4000和28000蛋白。与Ad7 Gomen株的相应区域相比，它们的核苷酸序列同源性分别为98.9%、97.3%、97.5%，推导的氨基酸序列同源性分别为96.6%、96.5%和96.9%。与Ad7 Grider株相比，它们的核苷酸序列同源性分别为100%、99.7%、99.7%，推导的氨基酸序列同源性分别为100%、99.1%和99.2%。