Nucleotide sequencing and generation of an infectious clone of adeno-associated virus 3
Nucleotide sequencing and generation of an infectious clone of adeno-associated virus 3
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DOI：
10.1006/viro.1996.0367
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发表时间：
1996-07-01
期刊：
影响因子：
3.7
通讯作者：
Brown, KE
中科院分区：
文献类型：
--
作者：
Muramatsu, S;Mizukami, H;Brown, KE
我们已经确定了腺相关病毒 3 (AAV-3) 的完整核苷酸序列并生成了感染性克隆。 AAV-3的单链DNA基因组长度为4726个核苷酸，正链包含两个大的开放阅读框；左侧开放阅读框编码非结构蛋白，右侧开放阅读框编码结构蛋白。编码区两侧是包含回文的相同反向末端重复序列，AAV-3 与自主细小病毒或红病毒几乎没有同源性，但与 AAV-2 具有 82% 的整体序列同源性。在氨基酸水平上，与 AAV-2 非结构 (Rep) 蛋白有 88% 同源性，与 AAV-2 衣壳蛋白有 87% 同源性。此外，AAV-3 与 AAV-2 的重要区别在于，在 p40 处缺乏典型的启动子序列（TATA 盒），并且在 p5 启动子的上游区域存在腺病毒相关转录因子 E4F 结合的共有序列。这些结果表明，AAV-3不仅由血清学上不同的结构蛋白组成，而且病毒传播也可能在转录水平上受到不同基因调控元件的控制。感染性克隆证实了该序列，可能有助于开发用于基因治疗的新载体。 (C) 1996 学术出版社
We have determined the complete nucleotide sequences of adeno-associated virus 3 (AAV-3) and generated an infectious clone. The single-stranded DNA genome of AAV-3 is 4726 nucleotides in length, The positive strand contains two large open reading frames; the left open reading frame encodes the nonstructural proteins and the right open reading frame encodes the structural proteins. The coding regions are flanked by identical inverted terminal repeat sequences containing palindromes, AAV-3 has little homology with the autonomous parvoviruses or erythroviruses but has 82% overall sequence homology with AAV-2. At the amino acid level there was 88% homology with AAV-2 nonstructural (Rep) proteins and 87% homology with AAV-2 capsid proteins. In addition, AAV-3 differed importantly from AAV-2 in the lack of a typical promoter sequence (TATA box) at p40 and the presence of the consensus sequence for adenovirus-related transcription factor E4F binding within the upstream region of the p5 promoter. These results suggest that AAV-3 not only consists of serologically distinct structural proteins but that viral propagation also may be controlled by different gene regulatory elements at the transcription level. The infectious clone confirmed the sequence and may be useful for developing new vectors for gene therapy. (C) 1996 Academic Press, Inc.