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嵌合逆转录病毒在体外和体内的适应性:具有扩展宿主范围的禽逆转录病毒载体的分离

Adaptation of chimeric retroviruses in vitro and in vivo: isolation of avian retroviral vectors with extended host range.

作者信息

Barsov E V, Payne W S, Hughes S H

机构信息

HIV Drug Resistance Program, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, Maryland 21702-1201, USA.

出版信息

J Virol. 2001 Jun;75(11):4973-83. doi: 10.1128/JVI.75.11.4973-4983.2001.

Abstract

We have designed and characterized two new replication-competent avian sarcoma/leukosis virus-based retroviral vectors with amphotropic and ecotropic host ranges. The amphotropic vector RCASBP-M2C(797-8), was obtained by passaging the chimeric retroviral vector RCASBP-M2C(4070A) (6) in chicken embryos. The ecotropic vector, RCASBP(Eco), was created by replacing the env-coding region in the retroviral vector RCASBP(A) with the env region from an ecotropic murine leukemia virus. It replicates efficiently in avian DFJ8 cells that express murine ecotropic receptor. For both vectors, permanent cell lines that produce viral stocks with titers of about 5 x 10(6) CFU/ml on mammalian cells can be easily established by passaging transfected avian cells. Some chimeric viruses, for example, RCASBP(Eco), replicate efficiently without modifications. For those chimeric viruses that do require modification, adaptation by passage in vitro or in vivo is a general strategy. This strategy has been used to prepare vectors with altered host range and could potentially be used to develop vectors that would be useful for targeted gene delivery.

摘要

我们设计并鉴定了两种具有双嗜性和嗜异性宿主范围的、新的具备复制能力的基于禽肉瘤/白血病病毒的逆转录病毒载体。双嗜性载体RCASBP-M2C(797-8)是通过在鸡胚中传代嵌合逆转录病毒载体RCASBP-M2C(4070A)(6)获得的。嗜异性载体RCASBP(Eco)是通过用嗜异性鼠白血病病毒的env区域替换逆转录病毒载体RCASBP(A)中的env编码区域而构建的。它在表达鼠嗜异性受体的禽DFJ8细胞中能高效复制。对于这两种载体,通过传代转染的禽细胞,可轻松建立在哺乳动物细胞上产生滴度约为5×10(6) CFU/ml病毒储备液的永久细胞系。一些嵌合病毒,例如RCASBP(Eco),无需修饰就能高效复制。对于那些确实需要修饰的嵌合病毒,在体外或体内传代进行适应性改造是一种常用策略。该策略已用于制备宿主范围改变的载体,并有可能用于开发对靶向基因递送有用的载体。

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