Morozov A Iu, Aleksandrova S S, Khodarev N N, Votrin I I
Mol Gen Mikrobiol Virusol. 1989 Apr(4):42-5.
The system of DNA recombination in vitro was constructed. It comprises two plasmids, the derivatives of pBR322 deleted in the genes for tetracycline resistance, and the recombinogenic extract of the thymus lymphocytes nuclei of mice. The system permits to study the effect of proteins and factors on the efficiency of recombination resulting in reconstruction of the tetracycline resistance gene. Double-strand cuts in one of the deleted plasmids were necessary for recombination. Double-strand cuts by Ca/Mg-dependent endonuclease of the human spleen lymphocytes nuclei were more efficient as compared with the ones of DNAase I, restriction endonucleases PaeI and SalI in the initiation of recombination. The possible role of Ca/Mg-dependent endonuclease in recombination in vivo is discussed.
构建了体外DNA重组系统。它由两个质粒组成，一个是pBR322在四环素抗性基因处缺失的衍生物，另一个是小鼠胸腺淋巴细胞细胞核的重组提取物。该系统可用于研究蛋白质和因子对重组效率的影响，重组可导致四环素抗性基因的重建。其中一个缺失质粒中的双链切割对于重组是必需的。与DNA酶I、限制性内切酶PaeI和SalI相比，人脾淋巴细胞细胞核的钙/镁依赖性内切酶进行的双链切割在重组起始方面更有效。讨论了钙/镁依赖性内切酶在体内重组中的可能作用。
