Sladek T L, Maniloff J
Department of Microbiology and Immunology, University of Rochester Medical Center, New York.
Isr J Med Sci. 1987 May;23(5):423-6.
Transfection of Acholeplasma laidlawii strain JA1 with viral DNAs that have been sequence-specifically methylated in vitro has led us to previously postulate that JA1 cells contain an enzyme which cleaves only DNA containing 5-methylcytosine, regardless of the nucleotide sequence containing that base. In this paper we show that an endonuclease activity is present in extracts from JA1 cells but not in extracts from a restriction-deficient variant of JA1. The partially purified enzyme cleaves not only DNA containing 5-methylcytosine, but also DNA containing no methylated bases. We discuss why this endonuclease activity may not have the same in vitro specificity as would be expected from in vivo experiments.
用在体外进行了序列特异性甲基化的病毒DNA转染莱氏无胆甾原体JA1菌株,这使我们之前推测JA1细胞含有一种酶,该酶仅切割含有5-甲基胞嘧啶的DNA,而不考虑含有该碱基的核苷酸序列。在本文中,我们表明JA1细胞提取物中存在一种核酸内切酶活性,而JA1的限制缺陷变体提取物中则不存在。部分纯化的酶不仅切割含有5-甲基胞嘧啶的DNA,还切割不含有甲基化碱基的DNA。我们讨论了为什么这种核酸内切酶活性在体外可能没有体内实验预期的相同特异性。