7型腺病毒疫苗株76.5～87mu核苷酸序列分析

Cloning and sequencing of adenovirus type 7 vaccine strain 76.5-87mu fragment

目的获得７型腺病毒疫苗株（Ａｄ７ｖ）７６５－８７ｍｕ核苷酸序列，分析该区段的基因结构和功能。方法从Ａｄ７疫苗株克隆出６８－８７ｍｕ核苷酸片段，应用Ｓａｎｇｅｒ双脱氧法进行核苷酸序列分析。结果Ａｄ７ｖ７６５～８７ｍｕ核苷酸序列长度为３５５７ｂｐ，推测此片段编码Ｅ３区１２１ｋＤ、１９２ｋＤ、２０１ｋＤ、２０５ｋＤ、１０３ｋＤ和１５２ｋＤ蛋白的一部分。所编码的６个蛋白与Ａｄ７原型株（Ａｄ７ｐ）和Ａｄ７ｈ（１９８６年以来南美流行的毒株）核苷酸高度同源（大于９７２％）。１６１ｋＤ蛋白由于缺失一个核苷酸，造成移码突变，编码提前终止。编码７７ｋＤ蛋白的ＯＲＦ由于缺失一段核苷酸而提前终止编码。

Adenovirus 7(Ad7)vaccine strain has been used more than 30 years in US army recruits for prevention of Ad7 infection and has been proven to be safe and effective.Recent years more attention has been paid to exploring the possibilities of using it for gene therapy or making recombinant vaccine.ln order to construct adenovirus 7vector, Ad 7 vaccine strain 76.5-87mu fragment was cloned and sequenced.The fragment contains 3 557 bp which encodes 6 proteins of E3 region,namely 12.1kD,19.2kD,20.1kD,20.5kD,10.3kD and partial of 15.2kD.Nucleotide sequences of all these proteins show a high homology with the corresponding region of human adenovirus prototype strain Ad 7(Ad 7p)and Ad 7h(the homology of nucleotide sequence of the 6 proteins is more than 97.2%).The equivalent of ORF 16.1kD in Ad7p and Ad7h was mutation due to a base pair deletion.The region corresponding to ORF 7.7kD in Ad7p was missing in Ad7v due to a deletion.The results should be helpful to the construction of Ad7v vectors.