Barcak G J, Tomb J F, Laufer C S, Smith H O
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
J Bacteriol. 1989 May;171(5):2451-7. doi: 10.1128/jb.171.5.2451-2457.1989.
Two Haemophilus influenzae Rd genes each complemented the pleiotropic defects of the recA-like mutation rec-1. One gene, fec, was isolated on a 3.6-kilobase-pair EcoRI restriction fragment by complementation of the Fec- phenotype of bacteriophage lambda. The other gene, rec, was identified on a 3.1-kilobase-pair EcoRI fragment by Southern hybridization by using recA-like gene probes from Erwinia carotovora and Pseudomonas aeruginosa PAO. In a rec-1 strain of H. influenzae, the cloned genes restored resistance to UV irradiation, transformation by chromosomal DNA, and spontaneous release of HP1 prophage to wild-type levels. The fec and rec genes were located on the cloned segments by insertion and deletion mutagenesis and subcloning. The restriction endonuclease cleavage maps of the two DNAs were similar but not identical. Southern hybridization demonstrated that the two EcoRI restriction fragments contained homologous DNA sequences, but a fec gene-specific probe was prepared. Each gene encoded a 38,000-dalton polypeptide.
两个流感嗜血杆菌Rd基因各自弥补了recA样突变rec-1的多效性缺陷。一个基因fec通过对噬菌体λ的Fec-表型进行互补,在一个3.6千碱基对的EcoRI限制片段上分离得到。另一个基因rec通过使用来自胡萝卜软腐欧文氏菌和铜绿假单胞菌PAO的recA样基因探针进行Southern杂交,在一个3.1千碱基对的EcoRI片段上得以鉴定。在流感嗜血杆菌的rec-1菌株中,克隆的基因将对紫外线照射的抗性、染色体DNA转化以及HP1原噬菌体的自发释放恢复到了野生型水平。通过插入和缺失诱变以及亚克隆,fec和rec基因被定位在克隆片段上。这两个DNA的限制性内切酶切割图谱相似但并不相同。Southern杂交表明这两个EcoRI限制片段包含同源DNA序列,但制备了一个fec基因特异性探针。每个基因编码一种38,000道尔顿的多肽。