NUCLEIC ACID FROM AN ADENO-ASSOCIATED VIRUS - CHEMICAL AND PHYSICAL STUDIES
NUCLEIC ACID FROM AN ADENO-ASSOCIATED VIRUS - CHEMICAL AND PHYSICAL STUDIES
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DOI：
10.1073/pnas.56.1.86
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发表时间：
1966-01-01
影响因子：
11.1
通讯作者：
SHATKIN, AJ
中科院分区：
文献类型：
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作者：
ROSE, JA;HOGGAN, MD;SHATKIN, AJ
据报道，基于吖啶橙子染色，在猿腺病毒15型制备物中发现的AAV含有双链DNA。1然而，AAV核酸尚未进一步表征。进行本工作以确定来自在人7型腺病毒株LL E46+（E46+）2，3的制备物中发现的AAV的核酸的化学和物理性质，并将这些性质与E46+ DNA的性质进行比较。结果表明，该AAV（现在命名为AAV血清型1、2）含有双链DNA，其碱基组成和分子量与E46+ DNA显著不同。材料：KB细胞系由Dr. M.绿色;人胚肾（HEK）细胞的原代单层培养物购自Flow Laboratories。光学级CsCl得自Harshaw Chemical Co.，来自Calbiochem的5 ′-脱氧核苷酸，来自Nuclear-Chicago和施瓦茨BioResearch，Inc.的胸苷-H3，来自新英格兰核公司的尿苷-H3，蛇毒磷酸二酯酶、经消化纯化的胰腺DNA酶I、2X结晶木瓜蛋白酶和结晶-冻干胰蛋白酶购自Worthington Biochemical Corp.。
AAV found in simian adenovirus type 15 preparations has been reported to contain double-stranded DNA on the basis of acridine orange staining. 1 However, AAV nucleic acid has not been further characterized. The present work was undertaken to define chemical and physical properties of nucleic acid from the AAV found in a preparation of human adenovirus type 7 strain LL E46+(E46+) 2, 3 and to compare these properties with those of E46+ DNA. The results indicate that this AAV, now designated AAV serotype 1, 2 contains double-stranded DNA with a base composition and molecular weight differing significantly from E46+ DNA.Experimental Procedure.-Materials: A KB cell line was provided by Dr. M. Green; primary monolayer cultures of human embryonic kidney (HEK) cells were purchased from Flow Laboratories. Optical grade CsCl was obtained from Harshaw Chemical Co., 5'-deoxynucleotides from Calbiochem, thymidine-H3 from Nuclear-Chicago and Schwarz BioResearch, Inc., uridine-H3 from New England Nuclear Corp., and P32-orthophosphate from International Chemical and Nuclear Corp. Snake venom phosphodiesterase, electrophoretically purified pancreatic DNase I, 2X crystallized papain, and crystallized-lyophilized trypsin were purchased from Worthington Biochemical Corp.