Gorbunova E E, Makarova N I, Shchelkunova G A
Vopr Virusol. 1980 May-Jun(3):360-4.
The possibility of using honeycomb moth larvae for titration of nuclear polyhedrosis virus (NPV) infectious DNA and determinations of transfection effectiveness was studied. Honeycomb moth larvae were shown to be a sensitive system for NPV DNA titration. DEAE-dextran used as a protector increased NPV DNA infectivity 1000-fold, LD50 in this instance being 2 X 10(8) molecules per larva. The method of NPV DNA infectivity determinations by the number of larvae with polyhedreae in the fatty tissue is more sensitive than infectivity determinations by the number of dead larvae and permits titrations of low DNA concentrations. The curve of DNA titration in the presence of DEAE-dextran by the number of larvae with polyhedrae in the fatty tissue allows to quantitate native DNA within the range of 0.01 to 5 micrograms/ml.
研究了使用巢蛾幼虫滴定核型多角体病毒(NPV)感染性DNA以及测定转染效率的可能性。结果表明,巢蛾幼虫是NPV DNA滴定的敏感系统。用作保护剂的DEAE-葡聚糖可使NPV DNA感染性提高1000倍,此时的半数致死剂量(LD50)为每只幼虫2×10⁸个分子。通过脂肪组织中多角体幼虫数量来测定NPV DNA感染性的方法比通过死亡幼虫数量测定感染性更为灵敏,并且能够滴定低浓度的DNA。利用脂肪组织中带有多角体的幼虫数量对存在DEAE-葡聚糖时的DNA进行滴定,所得曲线能够对0.01至5微克/毫升范围内的天然DNA进行定量。