Keseler I M, Kaiser D
Department of Biochemistry, Stanford University School of Medicine, CA 94305-5307, USA.
Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1979-84. doi: 10.1073/pnas.94.5.1979.
The rpoN gene encoding the transcription factor sigma54 in Myxococcus xanthus has been cloned using a heterologous rpoN probe. The sequence of the cross-hybridizing DNA confirmed the existence of an ORF 1518 bp long that encodes a well conserved member of the sigma54 family of sigma factors. Low- as well as high-stringency hybridizations detected only a single rpoN gene in the M. xanthus chromosome. In other bacteria, sigma54 is an alternative sigma, and null mutants are viable. However, all attempts to construct a strain containing a null mutation in the M. xanthus rpoN have been unsuccessful. Partial diploids of rpoN+/rpoN null are viable. Recombination experiments with such partial diploids showed the impossibility of constructing, either by segregation or by transduction, a viable null haploid under any of a wide range of growth conditions. The product of the rpoN gene, sigma54, therefore appears to be essential for growth in M. xanthus.
利用异源rpoN探针克隆了编码黄色粘球菌转录因子σ54的rpoN基因。交叉杂交DNA的序列证实存在一个1518 bp长的开放阅读框,该开放阅读框编码σ因子σ54家族中一个保守性良好的成员。低严谨度和高严谨度杂交在黄色粘球菌染色体中均仅检测到一个rpoN基因。在其他细菌中,σ54是一种替代σ因子,其缺失突变体是可存活的。然而,所有构建黄色粘球菌rpoN基因缺失突变体菌株的尝试均未成功。rpoN+/rpoN缺失的部分二倍体是可存活的。用这种部分二倍体进行的重组实验表明,在广泛的生长条件下,通过分离或转导构建一个可存活的缺失单倍体是不可能的。因此,rpoN基因的产物σ54似乎是黄色粘球菌生长所必需的。