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昆虫细胞系的建立在病毒学和昆虫学等领域的研究和应用中发挥着重要的作用。本研究由棉铃虫Helicoverpa armigera胚胎组织建立了两株细胞系,分别命名为QB-Ha-E-1和QB-Ha-E-5,在含10%胎牛血清的TNM-FH培养基中已传代60余代。两株细胞系均以圆形和短梭形细胞为主。DAF鉴定结果表明,两株细胞系均来源于棉铃虫胚胎,其扩增谱带与其他几种昆虫细胞系明显不同;QB-Ha-E-1和QB-Ha-E-5的第30代细胞群体倍增时间分别为63.7h和66.9h。两株细胞系均能被棉铃虫核型多角体病毒(HaSNPV)感染,4d的感染率分别为86.6%和56.5%,对甘蓝夜蛾Mamestra brassicae核型多角体病毒(MbNPV)7d的感染率均为15%左右,但对苜蓿银纹夜蛾Autographacalifornica核型多角体病毒(AcMNPV)侵染的反应不同。DAPI染色和基因组DNA电泳结果表明,AcMNPV可诱导QB-Ha-E-5细胞发生凋亡,极少数细胞内可形成多角体,但不能诱导QB-Ha-E-1细胞发生凋亡,其感染率为55.3%;两株细胞系均可被1.25μg/mL的放线菌素D诱导发生凋亡。两株细胞系具有相同的遗传背景,但对AcMNPV侵染的反应不同,可作为昆虫病毒和细胞之间相互关系以及细胞凋亡机制研究的理想材料。
The establishment of insect cell lines plays an important role in the research and application of virology and entomology. In this study, two cell lines were established from Helicoverpa armigera embryos, named as QB-Ha-E-1 and QB-Ha-E-5, respectively and cultured in TNM-FH medium containing 10% fetal bovine serum Passed more than 60 generations. Both cell lines were mainly round and short spindle cells. The results of DAF identification showed that both cell lines were derived from the embryos of H. armigera and their amplification bands were significantly different from those of other insect cell lines. The 30th generation of QB-Ha-E-1 and QB-Ha-E-5 Cell population doubling time was 63.7h and 66.9h respectively. Both cell lines were infected with HaSNPV, and the infection rates were 86.6% and 56.5% on the 4th day respectively. The infection rates of M. nipponense Mamestra brassicae nuclear polyhedrosis virus (MbNPV) for 7 days About 15%, but different responses to Autographa californica nuclear polyhedrosis virus (AcMNPV) infection of Autographa californica. The results of DAPI staining and genomic DNA electrophoresis showed that AcMNPV could induce the apoptosis of QB-Ha-E-5 cells and polyhedron could be formed in a few cells, but could not induce the apoptosis of QB-Ha-E-1 cells. The rate was 55.3%. Both cell lines could be induced apoptosis by 1.25μg / mL actinomycin D. Both cell lines have the same genetic background, but respond differently to AcMNPV infection and can be used as an ideal material for studying the relationship between insect viruses and cells and the mechanism of apoptosis.