| 禽腺联病毒全基因组的克隆及感染性病毒的拯救 |
| |
| 引用本文： | 王建业,孙怀昌,朱国强.禽腺联病毒全基因组的克隆及感染性病毒的拯救[J].中国预防兽医学报,2007,29(11):825-829. |
| |
| 作者姓名： | 王建业 孙怀昌 朱国强 |
| |
| 作者单位： | 扬州大学兽医学院,江苏,扬州,225009 |
| |
| 摘 要： | 为了克隆禽腺联病毒(Avian adeno-associated virus,AAAV)全基因组用于构建基因转移载体研究,以鸡胚致死孤儿病毒(CELO)作为辅助病毒与AAAV共接种SPF鸡胚进行AAAV的增殖,将AAAV约4.7kb双链基因组DNA与pCR2.1载体连接,构建了含AAAV全基因组的重组质粒pAAAV并进行了测序。序列分析表明,AAAV YZ-1株的基因组为4684bp,两端具有141bp的末端倒置重复序列和Rep蛋白结合位点特征序列,与GenBank中收录的AAAV DA-1株和VR-865株的核苷酸序列同源性分别为95.0%和92.2%。将pAAAV质粒转染CELO病毒感染的鸡胚肝细胞系,获得了感染性AAAV病毒粒子,结果证明克隆的AAAV基因组中存在与病毒复制和包装相关的正确关键序列,可用于重组AAAV载体的构建。
|
| 关 键 词： | 禽腺联病毒 基因组克隆 序列分析 病毒拯救 |
| 文章编号： | 1008-0589-（2007）11-0825-05 |
| 修稿时间： | 2006-11-28 |
Cloning of avian adeno-associated virus genome and rescue of the infectious virus |
| |
| WANG Jian-ye,SUN Huai-chang,ZHU Guo-qiang.Cloning of avian adeno-associated virus genome and rescue of the infectious virus[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(11):825-829. |
| |
| Authors: | WANG Jian-ye SUN Huai-chang ZHU Guo-qiang |
| |
| Institution: | College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China |
| |
| Abstract: | The genome of avian adeno-associated virus(AAAV)was cloned for construction of gene transfer vectors.AAAV was propagated in special pathogen-free chicken embryo using the chicken embryo lethal orphan(CELO)virus as the helper virus. The double-stranded genomic DNA was extracted from precipitated AAAV viron and cloned into pCR 2.1 vector.Sequence analysis showed that the genome of AAAV YZ-1 strain was 4 684 bp long with 141 bp inverted terminal repeats and typical Rep-binding element.The YZ-1 shared 95.0 % and 92.2 % nucleotide identity with the AAAV strains DA-1 and VR-865, respectively.The infectious virus was rescued by transfecting the recombinant plasmid pAAAV in chicken embryonic liver cell line in the presence of CELO virus.These data indicated that the recombinant plasmid pAAAV with essential elements involved in AAAV replication and packaging could be used for construction of recombinant AAAV vectors. |
| |
| Keywords: | avian adeno-associated virus genome cloning sequence analysis virus rescue |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
