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在果蝇 S2 细胞群体中对重组狂犬病病毒糖蛋白(RVGP)cDNA 转录和 mRNA 翻译的动力学研究。

Kinetic studies of recombinant rabies virus glycoprotein (RVGP) cDNA transcription and mRNA translation in Drosophila melanogaster S2 cell populations.

机构信息

Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, 05503-900, Brazil,

出版信息

Cytotechnology. 2013 Oct;65(5):829-38. doi: 10.1007/s10616-012-9522-6. Epub 2013 Jan 23.

Abstract

Recombinant rabies virus glycoprotein (RVGP) was expressed in cell membranes of stably transfected Drosophila S2 cells using constitutive and inducible promoters. Although with quantitative differences of RVGP expression in both systems, the cDNA transcription, as evaluated by relative RVGP mRNA levels measured by qRT-PCR, sustained the amount of RVGP producing cells and the RVGP volumetric (ΠRVGP) productivity. At the transition to the stationary cell growth phase, once the cell culture slowed down its rate of multiplication, an accumulation of RVGP mRNA and RVGP was clearly observed in both cell populations. Nevertheless, cell cultures performed under sub-optimal temperatures indicated that an envisaged increase in the RVGP production is not only dependent on cell growth rate, but essentially on optimal cell metabolic state.

摘要

使用组成型和诱导型启动子,在稳定转染的果蝇 S2 细胞的细胞膜中表达重组狂犬病病毒糖蛋白 (RVGP)。尽管两种系统中 RVGP 的表达存在定量差异,但通过 qRT-PCR 相对 RVGP mRNA 水平评估的 cDNA 转录维持了产生 RVGP 的细胞数量和 RVGP 的比生产率 (Π RVGP)。在过渡到静止细胞生长阶段时,一旦细胞培养降低其倍增速度,就可以在两种细胞群体中清楚地观察到 RVGP mRNA 和 RVGP 的积累。然而,在亚最佳温度下进行的细胞培养表明,预计增加 RVGP 的产量不仅取决于细胞生长速度,还主要取决于最佳细胞代谢状态。

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