Nouvel P, Panthier J J, Condamine H
Unité de Génétique des Mammifères, Institut Pasteur, Paris, France.
Virology. 1994 Oct;204(1):180-9. doi: 10.1006/viro.1994.1522.
A retroviral vector in which the gag and pol genes have been replaced by the NLS-lacZ reporter gene was derived from a cloned AKV-like virus. A complementing cell line expressing the gag and pol retroviral genes was constructed. The retroviral vector was demonstrated to replicate in the complementing cells. Since transfection is known to generate deletion variants of the introduced plasmid, we have examined whether it can give rise to viral forms with a replicating advantage over the initial vector. After transfection in complementing cells the spread of the vector was followed by X-gal staining. The fraction of stained cells increased for the first 10 days following transfection and was then stabilized to about 20% stained cells, thus defining two cell types; one with LacZ+ phenotype and one with LacZ- phenotype. Molecular analysis showed that the latter contains a deleted form of the virus preventing cell infection by the vector presumably through a mechanism of interference involving the viral env gene. Thus, interference results in the efficient block of vector expansion.
一种逆转录病毒载体，其gag和pol基因已被NLS-lacZ报告基因取代，该载体源自克隆的AKV样病毒。构建了表达gag和pol逆转录病毒基因的互补细胞系。已证明该逆转录病毒载体可在互补细胞中复制。由于已知转染会产生导入质粒的缺失变体，我们研究了它是否会产生比初始载体具有复制优势的病毒形式。在互补细胞中转染后，通过X-gal染色追踪载体的传播。转染后的前10天，染色细胞的比例增加，然后稳定在约20%的染色细胞，从而定义了两种细胞类型；一种具有LacZ+表型，另一种具有LacZ-表型。分子分析表明，后者含有病毒的缺失形式，但可能通过涉及病毒env基因的干扰机制来阻止载体感染细胞。因此，干扰导致载体扩增的有效阻断。
