AcMNPV突变株的蛋白表达时相研究

Time Course of Protein Expression in the Cell Infected with Mutants of AcMNPV

将苜蓿银纹夜蛾多核衣壳核型多角体病毒(Autograph Colifornica nuclear polyhedrosis virus,AcMNPV)的野生型株HR3和温度敏感突变株ts317、ts538、ts8感染草地贪夜蛾(Spodoptera frugiperda)Sf21细胞,并在允许温度(25℃)或非允许温度(33℃)下培养,分别采用过氧化物酶标记的抗P47蛋白、抗P143蛋白、抗多体蛋白和抗病毒结构蛋白的单克隆抗体检测病毒增殖过程各蛋白出现的时间。结果表明:1)P47蛋白是一种晚期(12hpi)表达蛋白,各突变株在允许温度(25℃)能够表达,但在非允许温度(33℃)不能表达。2)P143蛋白是一种早期(8hpi)表达蛋白,在允许温度和非允许温度时都能表达,ts8的表达量较少。3)在非允许温度条件下,蛋白质的合成速度高于允许温度。4)野生型和突变株ts317的病毒结构蛋白(P80、GP64、VP39、P24和PTP)在允许温度增殖下都能检测到,ts538和ts8表达量相对少些。5)除了GP64和P24外,ts538和ts8感染的细胞在非允许温度下不能表达病毒的结构蛋白。6)野生型毒株HR3在允许温度和非允许温度下的蛋白表达无明显差异。

In order to compare the difierence of protein expression in Autograph Californica nuclear polyhedrosis virus (AcMNPV) mutants, the Spodopterafrugiperda IPLB-SF-21 Cell was infected with wild type (HR3) and mutants (ts317, ts538, ts8), and incubated at permissive and nonpermissive temperature, then determined the time of protein expression with antibodies against P47, P143 and other viral structure proteins of budded virus.The results indicated; 1) P47 was expressed protein in late. The cells infected with mutants could be expressed the P47 at 25℃ and not at 33℃. 2) P143 was expressed protein early. The cells infected with mutants could be expressed the P143 at both temperature , but ts8 was the most weak. 3) Similar levels of structural proteins of AcMNPV were detected in both wild type and ts317 at 25℃, but ts538 and ts8 were slight weak than ts317-infected cell. Almost of BV proteins (except to GP64 and P2. 4) were not expressed in cells infected with ts538 and ts8 at 33℃.