Muskhelishvili G, Palm P, Zillig W
Max-Planck-Institut für Biochemie, Munich, FRG.
Mol Gen Genet. 1993 Mar;237(3):334-42. doi: 10.1007/BF00279436.
We present evidence for the existence of a conservative site-specific recombination system in Archaea by demonstrating integrative recombination of Sulfolobus shibatae virus SSV1 DNA with the host chromosome, catalysed by the SSV1-encoded integrase in vitro. The putative int gene of SSV1 was expressed in Escherichia coli yielding a protein of about 39 kDa. This protein alone efficiently recombined linear DNA substrates containing chromosomal (attA) and viral (attP) attachment sites; recombination with either negatively or positively supercoiled SSV1 DNA was less efficient. Intermolecular attA x attA and attP x attP recombination was also promoted by the SSV integrase. The invariant 44 bp "common attachment core" present in all att sites contained sufficient information to allow recombination, whilst the flanking sequences effected the efficiency. These features clearly distinguish the SSV1--encoded site--specific recombination system from others and make it suitable for the study of regulatory mechanisms of SSV1 genome--host chromosome interaction and investigations of the evolution of the recombination machinery.
我们通过证明柴田硫化叶菌病毒SSV1 DNA与宿主染色体的整合重组(由SSV1编码的整合酶在体外催化),为古菌中保守的位点特异性重组系统的存在提供了证据。SSV1的推定int基因在大肠杆菌中表达,产生一种约39 kDa的蛋白质。该蛋白质单独就能有效地重组含有染色体(attA)和病毒(attP)附着位点的线性DNA底物;与负超螺旋或正超螺旋的SSV1 DNA重组效率较低。SSV整合酶也促进分子间attA×attA和attP×attP重组。所有att位点中存在的不变的44 bp“共同附着核心”包含足以允许重组的信息,而侧翼序列影响重组效率。这些特征清楚地将SSV1编码的位点特异性重组系统与其他系统区分开来,使其适合于研究SSV1基因组与宿主染色体相互作用的调控机制以及重组机制的进化研究。