Guilliams T G, Teng M, Halligan B D
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226, USA.
Biochimie. 1997;79(1):13-22. doi: 10.1016/s0300-9084(97)87620-1.
We have previously identified a cDNA encoding part of the amino terminal portion of the large subunit of replication factor c (RF-C, AP-1), which we have named VDJP. Analysis of VDJP demonstrated that it has amino acid homology to bacterial DNA ligases and specific binding to the nonamer portion of the V(D)J recombination signal sequence motif. In this report, we demonstrate that VDJP is capable of forming a covalent bond between DNA fragments in a sequence dependent fashion. The VDJP mediated DNA ligation reaction is neither dependent on the presence of compatible DNA ends nor on sequence homology between the DNA fragments that are joined. Furthermore, we show that the covalent junction between the DNA fragments is resistant to proteases and phenol, and therefore not protein linked.
我们先前已鉴定出一种编码复制因子c(RF-C,AP-1)大亚基氨基末端部分的cDNA,我们将其命名为VDJP。对VDJP的分析表明,它与细菌DNA连接酶具有氨基酸同源性,并能与V(D)J重组信号序列基序的九聚体部分特异性结合。在本报告中,我们证明VDJP能够以序列依赖的方式在DNA片段之间形成共价键。VDJP介导的DNA连接反应既不依赖于兼容DNA末端的存在,也不依赖于连接的DNA片段之间的序列同源性。此外,我们表明DNA片段之间的共价连接对蛋白酶和苯酚具有抗性,因此不是蛋白质连接的。