Glebov O K, Abramian D S
Tsitologiia. 1985 Jun;27(6):678-87.
Using dot-hybridization with thymidine kinase gene (tk gene) of Herpes simplex virus type 1 (HSV 1) of DNA preparations obtained from isolated metaphase chromosomes and lysate fractions of metaphase cells, which presumably contain smaller particles compared to metaphase chromosomes, it has been shown that the tk gene of HSV 1 is localized in chromosomes of cells of transformant clones unstable in TK+-phenotype. The DNA isolated from the metaphase chromosomes from cells of transformant clones is 1.5- or 2-fold more efficient in transforming TK-Chinese hamster cells than is the total high molecular weight DNA from the same cells. Upon transformation of TK- cells by the high molecular weight DNA from the tk gene of HSV 1-containing clones, varying in the rate of the loss of TK+-phenotypes, the character "rate of the loss of transformant phenotype" is transferred together with the tk gene of HSV 1 in 22% of cases. Cells of rerevertant clones, produced from TK- subclones of transformant clones, display the rate of the loss of transformant phenotype characteristic of cells of parental TK+-clones. A comparison of the results allows a conclusion that DNA sequences, determining the character "rate of the loss of transformant phenotype", are linked tightly with the transforming DNA proper containing the tk gene of HSV 1, but are not localized inside such a DNA.
利用点杂交技术,对从分离的中期染色体以及中期细胞的裂解物组分中获得的DNA制剂进行检测,这些裂解物组分可能比中期染色体含有更小的颗粒,其中所用的DNA制剂来自1型单纯疱疹病毒(HSV 1)的胸苷激酶基因(tk基因)。结果表明,HSV 1的tk基因定位于TK +表型不稳定的转化克隆细胞的染色体中。从转化克隆细胞的中期染色体中分离出的DNA,在转化TK -中国仓鼠细胞方面,比来自相同细胞的总高分子量DNA效率高1.5倍或2倍。在用来自含HSV 1的tk基因的克隆的高分子量DNA转化TK -细胞时,这些克隆在TK +表型丧失速率上有所不同,在22%的情况下,“转化体表型丧失速率”这一特征与HSV 1的tk基因一起传递。由转化克隆的TK -亚克隆产生的回复克隆细胞,表现出亲本TK +克隆细胞特有的转化体表型丧失速率。对结果进行比较可以得出结论,决定“转化体表型丧失速率”这一特征的DNA序列与含有HSV 1的tk基因的转化DNA紧密相连,但并不定位于此类DNA内部。