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通过额外表达tax增加BLV感染细胞系中的抗原产生。

Increase of antigen production in BLV-infected cell lines via additional expression of tax.

作者信息

Wagner H J, Blankenstein P, Bondzio A, Ebner D, Risse S

机构信息

Free University Berlin, Department of Veterinary Medicine, Germany.

出版信息

Zentralbl Veterinarmed B. 1995 Nov;42(9):543-50. doi: 10.1111/j.1439-0450.1995.tb00746.x.

Abstract

The selection of animals infected with the bovine leukaemia virus (BLV) is performed by the immunological detection of antibodies against the virus, commonly using the antigen gp51. Furthermore, research is being carried out to develop protective vaccines against BLV that have gp51 as their main component. Taking both of these factors into account, it is clear that there will be an increasing requirement for the virus antigen gp51 for some time to come. The permanently BLV-infected foetal lamb kidney cell line FLK/BLV (and its sublines) has been proved to be the most useful culture for the mass production of the virus antigen. Stable cell lines producing higher quantities of BLV antigen have not been established, either by subcloning of the FLK/BLV or by infection of other permanent cells with BLV. Here, a report is made on efforts to increase the expression of gp51 in BLV-infected cells via the additional expression of homologous transactivating virus protein tax. Selectable tax expression vectors that integrate into the host cell genome were constructed using BL provirus DNA fragments. Highly productive FLK/BLV cells were transfected with these vectors. Following selection with G 418, gp51-producing cell lines were established and tested for their productivity for several months. Some tax-vector-containing cell lines have produced 1.5-2 times more gp51 than the highly productive parental control cell line FLK/BLV 44-1.

摘要

感染牛白血病病毒(BLV)的动物的选择是通过针对该病毒抗体的免疫检测来进行的,通常使用抗原gp51。此外,正在开展研究以开发以gp51为主要成分的针对BLV的保护性疫苗。考虑到这两个因素,很明显在未来一段时间内对病毒抗原gp51的需求将会增加。已证明永久感染BLV的胎羊肾细胞系FLK/BLV(及其亚系)是大规模生产病毒抗原最有用的培养物。无论是通过FLK/BLV的亚克隆还是用BLV感染其他永久细胞,都尚未建立产生更高量BLV抗原的稳定细胞系。在此,报告了通过额外表达同源反式激活病毒蛋白tax来增加BLV感染细胞中gp51表达的努力。使用BL前病毒DNA片段构建整合到宿主细胞基因组中的可选择tax表达载体。用这些载体转染高产FLK/BLV细胞。在用G418选择后,建立了产生gp51的细胞系并对其生产力进行了数月测试。一些含tax载体的细胞系产生的gp51比高产亲代对照细胞系FLK/BLV 44-1多1.5至2倍。

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