Otsuka H, Dubbs D R, Kit S
J Gen Virol. 1979 Feb;42(2):373-86. doi: 10.1099/0022-1317-42-2-373.
MKSA207 cells, a BALB/c mouse kidney line transformed by a tsA mutant of SV40, are temperature-dependent for the expression of the 'standard transformed phenotype'. At the permissive temperature (33.5 degrees C), the mKSA207 cells resembled wild-type (wt) SV40 transformants; they contained the intranuclear SV40 T antigen, grew to high saturation density in monolayer culture in either 10% or 0.5% serum, and also in methylcellulose suspension culture and became multinucleate in cytochalasin B. At the nonpermissive temperature (39.8 degrees C), the mKSA207 cells lost some of their transformed properties; they grew only to low density in 10% serum, hardly grew at all in 0.5% serum or in methylcellulose suspension culture, and remained mono- or binucleate in cytochalasin B. At 40 degrees C in low serum, mKSA207 cells lost the intranuclear T antigen and when fed 10% serum at 39.8 degrees C, accumulated large amounts of T antigen in the cytoplasm. Derivatives of mKSA207 have been selected at 39.8 degrees C in liquid medium and methylcellulose suspension culture. The heat adapted lines, like wt SV40 transformants, exhibited the standard transformed phenotype at both 33.5 and 39.8 degrees C. It is unlikely that acquisition of temperature-independence for the transformed phenotype was due to reversion of the tsA gene to wild-type because the heat-adapted cell lines displayed the cytoplasmic T antigen at 39.8 degrees C, characteristic of the parental mKSA207 cells and SV40 rescued from one of the heat-adapted lines was temperature sensitive for growth. The T antigen levels (complement fixation units per 10(6) cells) of heat-adapted lines grown at 39.8 degrees C were comparable to those of mKSA207 cells grown at 33.5 or 39.8 degrees C.
MKSA207细胞是由SV40的tsA突变体转化的BALB/c小鼠肾细胞系,其“标准转化表型”的表达依赖于温度。在允许温度(33.5℃)下,mKSA207细胞类似于野生型(wt)SV40转化体;它们含有核内SV40 T抗原,在10%或0.5%血清的单层培养中以及甲基纤维素悬浮培养中生长至高饱和密度,并在细胞松弛素B作用下形成多核细胞。在非允许温度(39.8℃)下,mKSA207细胞失去了一些转化特性;它们在10%血清中仅生长至低密度,在0.5%血清或甲基纤维素悬浮培养中几乎不生长,并且在细胞松弛素B作用下仍为单核或双核。在低血清中于40℃时,mKSA207细胞失去核内T抗原,而在39.8℃时给予10%血清时,细胞质中积累大量T抗原。已在39.8℃的液体培养基和甲基纤维素悬浮培养中筛选出mKSA207的衍生物。热适应细胞系与wt SV40转化体一样,在33.5℃和39.8℃时均表现出标准转化表型。转化表型获得温度独立性不太可能是由于tsA基因回复突变为野生型,因为热适应细胞系在39.8℃时显示出细胞质T抗原,这是亲本mKSA207细胞的特征,并且从其中一个热适应细胞系中拯救出的SV40对生长具有温度敏感性。在39.8℃生长的热适应细胞系的T抗原水平(每10(6)个细胞的补体结合单位)与在33.5℃或39.8℃生长的mKSA207细胞相当。