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Upregulated expression of the cDNA fragment possibly related to the virulence of Acanthamoeba culbertsoni

Authors
 Kyung-Il Im  ;  Kwang-Min Park  ;  Tai-Soon Yong  ;  Yong-Pyo Hong  ;  Tae-Eun Kim 
Citation
 Korean Journal of Parasitology, Vol.37(4) : 257-263, 1999 
Journal Title
 Korean Journal of Parasitology 
ISSN
 0023-4001 
Issue Date
1999
MeSH
Acanthamoeba/genetics* ; Acanthamoeba/pathogenicity* ; Amino Acid Sequence ; Animals ; Base Sequence ; DNA, Complementary/chemistry ; DNA, Complementary/genetics* ; Mice ; Mice, Inbred ICR ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Messenger/analysis ; RNA, Messenger/isolation & purification ; Virulence
Abstract
Identification of the genes responsible for the recovery of virulence in brain-passaged Acanthamoeba culbertsoni was attempted via mRNA differential display-polymerase chain reaction (mRNA DD-PCR) analysis. In order to identify the regulatory changes in transcription of the virulence related genes by the brain passages, mRNA DD-PCR was performed which enabled the display of differentially transcribed mRNAs after the brain passages. Through mRNA DD-PCR analysis. 96 brain-passaged amoeba specific amplicons were observed and were screened to identify the amplicons that failed to amplify in the non-brain-passaged amoeba mRNAs. Out of the 96 brain-passaged amoeba specific amplicons, 12 turned out to be amplified only from the brain-passaged amoeba mRNAs by DNA slot blot hybridization. The clone, A289C, amplified with an arbitrary primer of UBC #289 and the oligo dT11-C primer, revealed the highest homology (49.8%) to the amino acid sequences of UPD-galactose lipid transferase of Erwinia amylovora, which is known to act as an important virulence factor. The deduced amino acid sequences of an insert DNA in clone A289C were also revealed to be similar to cpsD, which is the essential gene for the expression of type III capsule in group B streptococcus. Upregulated expression of clone A289C was verified by RNA slot blot hybridization. Similar hydrophobicity values were also observed between A289C (at residues 47-66) and the AmsG gene of E. amylovora (at residues 286-305: transmembrane domains). This result suggested that the insert of clone A289C might play the same function as galactosyl transferase controlled by the AmsG gene in E. amylovora.
Files in This Item:
T199902462.pdf Download
DOI
10.3347/kjp.1999.37.4.257
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Environmental Medical Biology (환경의생물학교실) > 1. Journal Papers
Yonsei Authors
Yong, Tai Soon(용태순) ORCID logo https://orcid.org/0000-0002-3445-0769
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/174165
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