关键词: 烟草天蛾, 新细胞系, 重组蛋白, 生长曲线, 苜蓿银纹夜蛾核型多角体病毒

Abstract:

The development of new cell lines from unusual and non-pest species may provide important source for basic research and biotechnology application. In this study, three new insect cell lines designated as QB-Ms1-8, QB-Ms2-2, and QB-Ms2-7, respectively, were established from eggs of Manduca sexta. These cells were subcultured approximately 50 passages in TNMFH medium at 28℃. The majority of cells of the lines generally are spindle-shaped. The population doubling time of QB-Ms1-8, QB-Ms2-2 and QB-Ms2-7 cells was 51, 31 and 49 h, respectively. Although these lines were insusceptible to Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) with the infection rate of 33%-40% at 96 h post infection, QB-Ms2-2 cell line showed higher level of secreted SEAP production in comparison with BTI-Tn5B1-4 cell line. In this study, a SEAP high expression type cell line, QB-M2-2 was isolated from the three new cell lines established from M. sexta, which may provide a source for searching novel clones with higher production of recombinant proteins.

Key words: Manduca sexta, insect cell line, recombinant protein, growth curve, Autographa californica multiple nuclear polyhedrosis virus (AcMNPV)