Cloning and functional analysis of the sequences flanking mini-Tn5 in the magnetosomes deleted mutant NM4 of Magnetospirillum gryphiswaldense MSR-1  

作　　者：LI Feng1,2, LI Ying1, JIANG Wei1, WANG Zhenfang1 & LI Jilun1 1. State Key Laboratory for Agro-biotechnology and Department of Microbiology, China Agricultural University, Beijing 100094, China 2. Department of Biology, Huaibei Coal Industry Teachers’ College, Huaibei 235000, China 

出　　处：《Science China(Life Sciences)》2005年第6期574-584,共11页中国科学（生命科学英文版）

基　　金：This work was supported by the Chinese National Programs for High Technology Research and Development(Grant No.2001AA218041).

摘　　要：A magnetosome deleted mutant NM4 of Magnetospirillum gryphiswaldense MSR-1 was generated by mini-Tn5 transposon mutagenesis, and a 5045-bp fragment flanking mini-Tn5 in NM4 was cloned by Anchored PCR. Sequencing analysis showed that this fragment involved six putative open reading frames (ORFs); the mini-Tn5 was inserted into ORF4. Functional complementary test indicated that the 5045-bp fragment was required for biosynthesis of mag-netosomes in M. gryphiswaldense MSR-1. The protein encoded by ORF4 had 25% of identity with the chemotaxis protein CheYIII of Caulobacter crescentus CB15, and the protein encoded by ORF4 contained a conserved signal receiver domain that can receive the signal from the sensor partner of the bacterial two-component systems. It was suggested that the protein en-coded by ORF4 may take part in the signal transduction relating to biosynthesis of magneto-somes.A magnetosome deleted mutant NM4 of Magnetospirillum gryphiswaldense MSR-1 was generated by mini-Tn5 transposon mutagenesis, and a 5045-bp fragment flanking mini-Tn5 in NM4 was cloned by Anchored PCR. Sequencing analysis showed that this fragment involved six putative open reading frames (ORFs); the mini-Tn5 was inserted into ORF4. Functional complementary test indicated that the 5045-bp fragment was required for biosynthesis of mag-netosomes in M. gryphiswaldense MSR-1. The protein encoded by ORF4 had 25% of identity with the chemotaxis protein CheYIII of Caulobacter crescentus CB15, and the protein encoded by ORF4 contained a conserved signal receiver domain that can receive the signal from the sensor partner of the bacterial two-component systems. It was suggested that the protein en-coded by ORF4 may take part in the signal transduction relating to biosynthesis of magneto-somes.

关 键 词：MAGNETOSPIRILLUM gryphiswaldense  MAGNETOSOME deleted mutant  gene cloning  functional analysis. 

分 类 号：Q78[生物学—分子生物学]