Mertz J E
Virology. 1984 Jan 15;132(1):173-85. doi: 10.1016/0042-6822(84)90101-6.
A simple mixed-plaque assay procedure for determining complementation between pairs of SV40 mutants is described. Since the data obtained can be quantified with respect to both the relative numbers and sizes of plaques, this method is less likely than other complementation assay procedures to yield false-positive results and gives some indication as to the efficiency of the complementation. This quantitative assay procedure, which works with both viral DNA and virions, was used to examine the complementation properties of a variety of late region temperature-sensitive and deletion mutants of SV40. Conclusions reached from these studies included (i) the complementation observed between tsB and tsC mutants is definitely intragenic; (ii) D mutants define a true complementation group; (iii) sequences within the last 68 carboxyl-terminal amino acids of VP-3 are necessary for expression of the D function; and (iv) when linked to the last 19 carboxyl-terminal amino acids of VP-1, the first 136 amino-terminal amino acids of VP-2 are sufficient for expression of the E function. Lastly, the collection of deletion mutants described here may be useful in identifying both structural and regulatory functions of the virion proteins and in analyzing some aspects of viral mRNA biogenesis.
本文描述了一种用于确定SV40突变体对之间互补作用的简单混合噬菌斑测定程序。由于所获得的数据可以在噬菌斑的相对数量和大小方面进行量化，因此该方法比其他互补测定程序产生假阳性结果的可能性更小，并且能给出一些关于互补效率的指示。这种适用于病毒DNA和病毒粒子的定量测定程序，被用于检测多种SV40晚期区域温度敏感型和缺失型突变体的互补特性。这些研究得出的结论包括：（i）tsB和tsC突变体之间观察到的互补作用肯定是基因内的；（ii）D突变体定义了一个真正的互补组；（iii）VP-3最后68个羧基末端氨基酸内的序列对于D功能的表达是必需的；（iv）当与VP-1的最后19个羧基末端氨基酸相连时，VP-2的前136个氨基末端氨基酸足以表达E功能。最后，本文所述的缺失突变体集合可能有助于鉴定病毒粒子蛋白的结构和调节功能，以及分析病毒mRNA生物合成的某些方面。
