Matsuura M, Noguchi T, Yamaguchi D, Aida T, Asayama M, Takahashi H, Shirai M
Division of Biotechnology, School of Agriculture, Ibaraki University, Japan.
J Bacteriol. 1996 Jun;178(11):3374-6. doi: 10.1128/jb.178.11.3374-3376.1996.
The sre gene (ORF469) of the R4 phage encodes a protein similar to the resolvase-DNA invertase family proteins. Insertional gene disruption of sre prevented a lysogen from entering the lytic cycle, implying that Sre protein is a site-specific recombinase needed for excision of the R4 prophage genome (M. Matsuura, T. Noguchi, T. Aida, M. Asayama, H. Takahashi, and M. Shirai, J. Gen. Appl. Microbiol. 41:53-61, 1995). To determine whether this sre gene is also necessary for the integration reaction, we studied its function by integration plasmid analysis. When deletions, frameshifts, and site-directed mutations that caused an amino acid substitution of Ser-17 for Ala were introduced into the sre structural gene, transformation efficiency of Streptomyces parvulus 2297 with these plasmid DNAs was severely reduced. However, an adenine insertion just before the possible initiation codon of the sre gene did not significantly decrease the efficiency. These data suggest that the Sre protein is a site-specific recombinase responsible for integration of the R4 phage genome.
R4噬菌体的sre基因(ORF469)编码一种与解离酶- DNA 转化酶家族蛋白相似的蛋白质。sre基因的插入性基因破坏阻止了溶原菌进入裂解周期,这意味着Sre蛋白是R4原噬菌体基因组切除所需的位点特异性重组酶(M. Matsuura、T. Noguchi、T. Aida、M. Asayama、H. Takahashi和M. Shirai,《应用微生物学杂志》41:53 - 61,1995年)。为了确定该sre基因对于整合反应是否也是必需的,我们通过整合质粒分析研究了它的功能。当将导致Ser - 17被Ala氨基酸取代的缺失、移码突变和定点突变引入sre结构基因时,这些质粒DNA对小链霉菌2297的转化效率严重降低。然而,在sre基因可能的起始密码子之前插入一个腺嘌呤并没有显著降低效率。这些数据表明Sre蛋白是负责R4噬菌体基因组整合的位点特异性重组酶。