Development of porcine adenovirus-3 as an expression vector
Development of porcine adenovirus-3 as an expression vector
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DOI：
10.1099/0022-1317-80-3-563
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发表时间：
1999-03-01
影响因子：
3.8
通讯作者：
Babiuk, LA
中科院分区：
文献类型：
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作者：
Reddy, PS;Idamakanti, N;Babiuk, LA
猪腺病毒3型（Porcine adenovirus-3，PAV-3）是在大肠杆菌BJ 5183中通过同源重组获得的一种表达载体。当Pad限制并转染到猪睾丸细胞中时，质粒产生感染性病毒。通过构建几种PAV-3重组体证明了该方法的潜力。E3区的一部分，这是不必要的病毒复制在细胞培养条件下，被确定和删除的病毒基因组。从猪伪狂犬病病毒（pseudorabiesvirus，PRV）E3区克隆并表达了能在猪体内产生中和抗体的PRV糖蛋白D（gD）基因。在另一个实验中，右反向末端重复序列和E4区的启动子之间的区域用于克隆和表达在SV 40立即早期启动子控制下的氯霉素乙酰转移酶（CAT）基因。观察到CAT基因的表达与CAT基因的方向无关。这些结果表明，辅助病毒非依赖性重组PAV-3可用作表达载体，并具有作为重组疫苗载体在猪中的潜力。
Porcine adenovirus-3 (PAV-3) was developed as an expression vector using homologous recombination in Escherichia coli BJ 5183, As a prerequisite, the complete genome of PAV-3 was first introduced as a Pad restriction fragment into a bacterial plasmid. The plasmid, when Pad restricted and transfected into swine testicular cells, produces an infectious virus. The potential of this procedure was demonstrated by the construction of several PAV-3 recombinants. Part of the E3 region, which is nonessential for virus replication under cell culture conditions, was identified and deleted from the virus genome. The gene for glycoprotein D (gD) of pseudorabies virus (PRV), which elicits PRV-neutralizing antibodies in pigs, was cloned and expressed from the E3 region of PAV-3, A 50 kDa polypeptide was identified in recombinant PAV-3-infected cell lysates by immunoprecipitation assays using go-specific monoclonal antibodies. In another experiment, a region between the right inverted terminal repeat and the promoter of the E4 region was used to clone and express the chloramphenicol acetyltransferase (CAT) gene under the control of SV40 immediate early promoter. CAT gene expression was observed irrespective of the orientation of the CAT gene. These results indicate that the helper-independent recombinant PAV-3 could be used as an expression vector and has potential as a recombinant vaccine vector in pigs.