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Journal Article
表达大熊猫源犬瘟热病毒H蛋白复制缺陷型重组腺病毒的构建及鉴定
2021
Overview
为构建能够表达大熊猫源犬瘟热病毒（CDV）分离株giant panda/SX/2014 H蛋白的复制缺陷型重组腺病毒，利用反转录聚合酶链反应（RTPCR）扩增株giant panda/SX/2014 H基因，通过TOPO连接克隆至入门载体pENTR/DTOPO，得到重组入门质粒pENTR/DTOPOCDVH，然后与腺病毒骨架载体pAd/CMV/V5DEST进行LR重组，获得重组质粒pAdCDVH；重组质粒用PacⅠ酶切线性化后转染293A细胞，获得重组腺病毒rAdCDVH，连续传代后对获得的重组腺病毒分别进行病毒滴定、免疫荧光和Western blot鉴定。经免疫荧光方法及Western blot鉴定结果表明，重组腺病毒rAdCDVH感染293A细胞、Vero细胞后均可以成功表达H蛋白，获得的重组腺病毒的滴度高达1010IFU/mL。成功获得能够表达大熊猫源CDV H蛋白的重组腺病毒rAdCDVH，在进一步证明其安全有效后，可为大熊猫犬瘟热免疫预防提供候选疫苗株。
Publisher
Editorial Department of Chinese Journal of Wildlife
Subject
