Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Homologous Expression and T3SS-Dependent Secretion of TAP-Tagged Xo2276 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract and Its Direct In Vitro Recognition of Putative Target DNA Sequence

  • Kim, Seunghwan (Department of Advanced Technology Fusion, Konkuk University) ;
  • Nguyen, Thi-Dieu-Hanh (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Lee, Joohee (Department of Green Life Science, College of Convergence, Sangmyung University) ;
  • Hong, Myoung-Ki (Department of Advanced Technology Fusion, Konkuk University) ;
  • Pham, Tan-Viet (Department of Advanced Technology Fusion, Konkuk University) ;
  • Ahn, Yeh-Jin (Department of Green Life Science, College of Convergence, Sangmyung University) ;
  • Lee, Byoung-Moo (Genomics Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Han, Ye Sun (Department of Advanced Technology Fusion, Konkuk University) ;
  • Kim, Dong-Eun (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Kim, Jeong-Gu (Genomics Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Kang, Lin-Woo (Department of Biological Sciences, Konkuk University)
  • Received : 2012.07.23
  • Accepted : 2012.08.30
  • Published : 2013.01.28
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Abstract

Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoosusceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAPtagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.

Keywords

References

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