Comparison of gene diversity in Riemerella anatipestifer
Date Issued
2006
Date
2006
Author(s)
DOI
zh-TW
Abstract
Abstract
Riemerella anatipestifer infection is an economic important disease in waterfowl and turkey industries. Twenty-one different serovars of R. anatipestifer have been reported and serovar 2 is the major type in Taiwan. PCR-based subtractive hybridization was employed to isolate DNA fragments in a tested R. anatipestifer strain. Serovar 2 of Taiwan local strain presented the specific fragments but was absent in reference avirulent vaccine strain. Thirty six specific DNA fragments in the local wild strain were obtained. The fragments were sequenced and their DNA sequences were analyzed by BLAST program. Among them 9 DNA fragments (genes) related to R. anatipestifer plasmids, 2 DNA fragments related to B. fragilis genes, 1 DNA fragment related to Yersinia, 1 DNA fragment related to eubacterium partial 16S rRNA gene and the other 23 DNA fragment had no DNA sequence homologs in the Genbank. Protein analysis showed that 5 clones had homology to hypothetic proteins, 9 clones were similar to negative regulator of β-lactamase, replicase, uroporphyrinogen decarboxylase, integrase, lysozyme, replication protein, ATPase, and addiction module toxin, respectively and 16 clones had a low similarity to bacterium protein. The other 6 clones encode proteins with unknown function. Subtractive hybridization technique is successfully identifying genes in field isolated R. anatipestifer that are absent in the reference avirulent vaccine strain. These results provide new insights into R. anatipestifer genetic diversity. Attempts to generate mutants by using transposons and shuttle vectors, but failed. The reasons that we are unable to generate mutants in R. anatipestifer with the broad-host-range plasmids and transposons are: (1). the plasmids and transposons were unable to transfer into the R. anatipestifer. (2). the plasmids and transposons might be degraded by R. anatipestifer restriction system. (3). the antibiotic gene from the plasmids or transposons could not express in R. anatipestifer and (4). The promoter of the transposase gene (tnp) could not be recognized by R. anatipestifer’s RNA polymerase and no transposase was expressed.
Subjects
鴨疫里莫氏桿菌
基因體相減雜交
跳躍子
穿梭載體
Riemerella anatipestifer
Genome subtractive hybridization
Transposon
Shuttle vector
SDGs
Type
thesis
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