1992 Fiscal Year Final Research Report Summary
MOLECULAR ANALYSIS OF THE GENES ENCODING CYSTEINE PROTEASES FROM PARASITIC PROTOZOA
| Project/Area Number |
03670202
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
寄生虫学(含医用動物学）
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| Research Institution | TOKAI UNIVERSITY |
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Principal Investigator |
KANEDA Yoshimasa DEPARTMENT OF INFECTIOUS DISEASES, TOKAI UNIVERSITY SCHOOL OF MEDICINE, PROFESSOR, 医学部感染症学, 教授 (60051471)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Tomoo DEPARTMENT OF INFECTIOUS DISEASES, TOKAI UNIVERSITY SCHOOL OF MEDICINE, INSTRUCT, 医学部感染症学, 助手 (50192175)
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| Project Period (FY) |
1991 – 1992
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| Keywords | Trypanosoma cruzi / Trypanosoma rangeli / Cysteine protease / Invasion to the cells / Pathogenicity / DNA mismatch repair gene / 病原性 |
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| Research Abstract |
Recent observations have indicated that the cysteine protease of parasitic protozoa is linked to invasion of host cells. It is of interest, therefore, to investigate the molecular properties of the enzymes of both invasive and non-invasive trypanosomes. We isolated the gene fragments encoding cysteine proteinase from several trypanosomes, using the polymerase chain reaction, and analyzed the amino acid sequences of the DNA fragments encoding the active site.
We also compared the amino acid sequences of the active sites among several strains of Trypanosoma cruzi, but little differences in the sequences were observed. Furthermore, two fragments were obtained from genomic DNA of non-pathogenic T. rangeli. The amino acid sequence of one fragment showed good homology with those of the active sites of known cysteine proteinases. The other fragment had a homologous sequence with the DNA mismatch repair gene from the yeast, Saccharomyces cerevisiae. Northern blot analysis revealed that both gene fragments were specifically expressed in T. rangeli as 1.7 Kb mRNA. The karyotype nap of the chromosomes was obtained from pulse-field gradient gel electrophoresis and Southern blot hybridization with these genes. Both fragments hybridized with the same two chromosomes.
Our goals are to further elucidate the properties and functions of this important enzyme, which we hypothesize is involved in the parasite's invasive ability.
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