Jiang Xiu-li, Wang Jian-wei, Wen Le-ying, Shi Chang-xin, Hong Tao
Department of Diarrhea Viruses Institute for Viral Disease Control and Prevention, Chinese Center For Disease Control and Prevention, Beijing 100052, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2003 Dec;17(4):305-9.
To complete the full-length sequencing of the human adenovirus type 7 vaccine strain (Ad7v) for novel vector constructing.
The Ad7v DNA was digested with SalI and the 17.5-68.0 map unit (mu) fragment was cloned and sequenced. The homology of encoding sequence of Ad7v hexon to those of group A,C,D,E,F and other numbers of group B was accomplished with the software CLUSTAL.V. The three-dimensional structure of the Ad7v hexon was predicted with the RasMo12.71.
The fragment contains 17,596 bp, part of E2 and late gene L1, L2 and L3 were encoded by this region. Polypeptide encoded by hexon gene lies in L3 region, which is composed of 934 amino acids. Multiple sequence alignment with the other nine known hexon protein sequences suggested that the variable sequences are mainly concentrated on seven regions, namely hypervariable regions (HVRs). The seven HVRs are related to type-specificity and group-specificity. The three-dimensional structure of the Ad7v hexon revealed that the variable regions are located in the I1 and I2 loops of the molecule mostly on the tower of the hexon.
The full-length genome sequencing of Ad7v was accomplished at last. Since the deduced amino acid sequence of Ad7v hexon was quite different from other adenoviral vectors such as Ad5 and Ad2, this virus can be potentially used for the construction of novel gene delivery vectors to counterpart the immunity to the vectors widely used at present.
完成人7型腺病毒疫苗株(Ad7v)的全长测序,用于构建新型载体。
用SalI酶切Ad7v DNA,克隆并测序17.5 - 68.0图谱单位(mu)的片段。使用CLUSTAL.V软件完成Ad7v六邻体编码序列与A、C、D、E、F组及其他B组腺病毒编码序列的同源性分析。用RasMo12.71软件预测Ad7v六邻体的三维结构。
该片段包含17596 bp,此区域编码部分E2基因及晚期基因L1、L2和L3。六邻体基因编码的多肽位于L3区域,由934个氨基酸组成。与其他9种已知六邻体蛋白序列的多重序列比对表明,可变序列主要集中在7个区域,即高变区(HVRs)。这7个HVRs与型特异性和组特异性相关。Ad7v六邻体的三维结构显示,可变区大多位于六邻体分子塔的I1和I2环上。
最终完成了Ad7v的全基因组测序。由于Ad7v六邻体推导的氨基酸序列与Ad5和Ad2等其他腺病毒载体有很大差异,该病毒有望用于构建新型基因递送载体,以应对目前广泛使用的载体的免疫问题。