Adolph K W, Butler P J
Philos Trans R Soc Lond B Biol Sci. 1976 Nov 30;276(943):113-22. doi: 10.1098/rstb.1976.0102.
The conditions previously reported as necessary for the reassembly of spherical viruses have been distinctly unphysiological and such reassembly cannot be related directly to the in vivo reaction. Mild conditions for the in vitro reassembly of cowpea chlorotic mottle virus (CCMV) from its isolated components have now been described (Adolph & Butler 1975) and the reassembled virus characterized. This reassembly involved the co-aggregation of the RNA and protein around neutrality and at ionic strength 0.2, giving yields of 70% encapsidation at pH 6.0. The reaction was independent of temperature over the range 5-25 degrees C and did not require the presence of Mg2+ ions. The reassembled virus shows a stability similar to that of native CCMV, with the same change in sedimentation coefficient around pH 6.5. The molecular mass and buoyant density in CsCl are also the same as those of native CCMV, while the electron microscope reveals a surface morphology on the reassembled particles like that on native CCMV. Analysis of the number-average, mass-average, and Z-average molecular masses of the purified protein at both pH 6.0 and pH 7.5 suggests that the active unit for reassembly is a dimer of the protein subunit.
先前报道的球状病毒重新组装所需的条件明显不符合生理状态,且这种重新组装无法直接与体内反应相关联。现在已经描述了从其分离的组分在温和条件下体外重新组装豇豆花叶病毒(CCMV)的方法(阿道夫和巴特勒,1975年),并对重新组装的病毒进行了表征。这种重新组装涉及RNA和蛋白质在接近中性和离子强度为0.2的条件下共同聚集,在pH 6.0时包封率达70%。该反应在5至25摄氏度范围内与温度无关,且不需要Mg2+离子的存在。重新组装的病毒显示出与天然CCMV相似的稳定性,在pH 6.5左右沉降系数有相同变化。在CsCl中的分子量和浮力密度也与天然CCMV相同,而电子显微镜显示重新组装颗粒的表面形态与天然CCMV的相似。对纯化蛋白质在pH 6.0和pH 7.5时的数均分子量、质均分子量和Z均分子量的分析表明,重新组装的活性单位是蛋白质亚基的二聚体。