粉纹夜蛾细胞系QB-Tn9-4s的克隆以及克隆株生物学特性的研究

Cloning of Trichoplusia ni cell line QB-Tn9-4s and characterization of its clones

对粉纹夜蛾Trichoplusiani细胞系QB-Tn9-4s进行细胞克隆,获得了8个细胞克隆株,分别命名为QB-Tn-A、B、C、D、E、F、G和H。对基因组DNA进行RAPD-PCR鉴定,各细胞克隆株与原始细胞系具有相同的DNA扩增谱带。各细胞克隆株在形态和生物学特性方面表现出一定的差异。克隆株QB-Tn-A、B、C、D和E以梭形细胞为主,大约占细胞总数的60%～80%;F、G和H以棒状细胞为主,比例分别为44.5%、49.5%和80.0%。8个克隆株对苜蓿银纹夜蛾核型多角体病毒(AcMNPV)均较敏感,感染率均在92%以上,平均每个细胞病毒多角体(OBs)产量在78～110个之间,其中克隆株QB-Tn-A多角体产量最高达110个,略高于BTI-Tn5B1-4和QB-Tn9-4s,明显高于Sf-9细胞;克隆株QB-Tn-E、H、A和C的出芽性病毒(BV)产量与原始细胞系(3.37×107TCID50/mL)接近,而其它4株均低于原始细胞系。

It is well known that the characteristics among cell lines from embryo are different. In this paper, we described clone and isolation of culture cell from low passage Trichoplusia ni cell line QB-Tn9-4s and reported their growth characteristics and production of occlusion bodies （OBs）. Eight cell clones were successfully derived from the established cell line and designated QB-Tn-A, B, C, D, E, F, G and H, respectively. RAPD-PCR analysis of the genomic DNA of the eight clones confirmed their genetic identity as QB-Tn9-4s. The morphology and biological characterization of the cell clones were different from each other. Clones QB-Tn-A, B, C, D and E consist of approximately 60% - 80% spindle-shaped cells, clones F, G and H were mostly made up of 44. 5% , 49.5% and 80. 0% comma-shaped cells, respectively. Eight clones were highly susceptible to AcMNPV virus with an infection of over 92% cells with production of around 78 - 110 OBs per cell. The clone QB-Tn-A produced the highest number of OBs per cell when compared with BTI-Tn5B1-4, QB-Tn9-4s and Sf-9. The productions of AcMNPV buded virus （BV） of all clones tended to different from each other, the clones QB-Tn-E, H, A, and C were similar to the primary cell line QB-Tn9-4s （3.37 × 10^7 TCID50/mL） , and others were lower than the primary cell line.