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KMID : 0545120000100020161
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Journal of Microbiology and Biotechnology
2000 Volume.10 No. 2 p.161 ~ p.168
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Cloning and Sequencing of the Gene Involved in Morphological Chang of Zoogloea ramigera 115SLR
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Lee, Sam Pin
Kim, Tae Rahk/Anthony John Sinskey
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Abstract
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Plasmid pLEX3 isolated from the recombinant cosmid library of Zoogloea ramigera 115 was found to be responsible for the restoration of the rugose colony phenotype. To confirm the essential region responsible for the complementation, subclones were constructed from plasmid pLEX3 and transformed into mutant strain Z ramigera 115SLR. The recombinant plasmids pLEX10 and pLEX11 were shown to complement the slime-forming property of Z. ramigera 115SLR. In a compositional analysis of the exopolysaccharides from Z ramigera 115, Z ramigera 115SLR, and Z ramigera 115SLR harboring plasmid pLEX11, the exopolysaccharides showed a similar composition with glucose, galactose, and side chain groups. The complete nucleotide sequence of the 3.25kb genomic DNA insert in plasmid pLEX11 was determined and its analysis identified two open reading frames which could encode two proteins. The gene products derived from the two open reading frames were confirmed by an in vivo transcription using a T7-RNA polymerase. The ORF1 produced a 30kDa protein, whereas the ORF2 was found responsible for the complementation of the morphological mutation and produced a 14kDa protein. An in vivo gene expression of plasmid pTEX10 showed another open reading frame encoding a 50kDa protein. The gene products from ORF1 and ORF2 are regarded as novel proteins which do not show any homology with other proteins.
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