AcMNPV突变株的蛋白表达时相研究

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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。 Wild-type strain HR3 and temperature sensitive mutants ts317, ts538 and ts8 of Autograph Coliwifornica nuclear polyhedrosis virus (AcMNPV) were infected with Sf21 cells of Spodoptera frugiperda, And cultured at the allowable temperature (25 ° C) or non-allowable temperature (33 ° C). Peroxidase-labeled anti-P47 protein, anti-P143 protein, anti-polytopic protein and anti-viral structural protein monoclonal antibody were used to detect the virus Proliferation process of each protein appears time. The results showed that: 1) P47 protein is a late expression protein (12hpi), and each mutant can express at the allowable temperature (25 ℃), but can not express at the non-allowable temperature (33 ℃). 2) The P143 protein is an early (8hpi) expressed protein that is expressed at both tolerated and unallowed temperatures, with less expression of ts8. 3) Under non-allowable temperature conditions, the rate of protein synthesis is higher than the allowable temperature. 4) The viral structural proteins of ts317 (P80, GP64, VP39, P24 and PTP) of wild-type and mutant ts317 were detected under temperature-tolerant conditions, and ts538 and ts8 were relatively less expressed. 5) In addition to GP64 and P24, ts538 and ts8 infected cells could not express viral structural proteins at non-permissive temperatures. 6) The wild-type strain HR3 showed no significant difference in protein expression between the allowable temperature and the non-allowable temperature.
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