Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Isolation and Characterization of vasa Gene of Triploid and Diploid Human Lung Flukes (Paragonimus westermani)

폐흡충의 이배체와 삼배체 vasa 유전자 분석 및 특징

  • Lee, Keun-Hee (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Yu, Hak-Sun (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Hur, Jae-Won (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Yu, Sung-Suk (Department of Life Science Gwangju Institute of Science and Technology) ;
  • Choi, Sun-Hee (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Park, Sang-Kyun (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Lee, Sun-Joo (Department of Parasitology, School of Medicine, Pusan National University) ;
  • Chung, Dong-Il (Department of Parasitology, School of Medicine, Kyungpook National University) ;
  • Kong, Hyun-Hee (Department of Parasitology, School of Medicine, Kyungpook National University) ;
  • Ock, Mee-Sun (Department of Parasitology, Kosin University College of Medicine) ;
  • Jeong, Hae-Jin (Department of Parasitology, School of Medicine, Pusan National University)
  • 이근희 (부산대학교 의과대학 기생충학 교실) ;
  • 유학선 (부산대학교 의과대학 기생충학 교실) ;
  • 허재원 (부산대학교 의과대학 기생충학 교실) ;
  • 유성숙 (광주과학기술연구소) ;
  • 최선희 (부산대학교 의과대학 기생충학 교실) ;
  • 박상균 (부산대학교 의과대학 기생충학 교실) ;
  • 이선주 (부산대학교 의과대학 기생충학 교실) ;
  • 정동일 (경북대학교 의과대학 기생충학 교실) ;
  • 공현희 (경북대학교 의과대학 기생충학 교실) ;
  • 옥미선 (고신대학교 의과대학 기생충학 교실) ;
  • 정해진 (부산대학교 의과대학 기생충학 교실)
  • Published : 2007.04.25
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Abstract

In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.

폐흡충은 국내를 비롯한 아시아에서 폐흡충증을 일으키는 중요한 기생충이다. 이러한 폐흡충은 이배체 와 삼배체가 국내에 알려져 있다. 이배체 폐흡충은 양성생식을 하고 삼배체 폐흡충은 단위생식을 하는 것으로 알려져 있다. 그러나 이러한 원인에 대하여서는 알려진 바가 없다. 생식유전자 중 초파리에서 최초 분리되었고 포유동물에서도 그 기능이 밝혀진 vasa 유전자가 가장 유명하다. 이 유전자는 생식세포의 분화에 관여하며 종에 따라서는 정자생성에도 관여하는 것으로 알려져 있다. 이번 연구는 삼배체 폐흡충의 단위생식과 vasa 유전자와의 관계를 규명해 보고자 하였다. 폐흡충의 이배체와 삼배체 성충으로부터 vasa 유전자 전체의 염기서열을 얻을 수 있었다. 두 염기서열의 경우 8개의 ATP-binding domain이 관찰되었고 helicase가 결합할 것으로 예상되는 RGG motif도 관찰되었다. 총 622아미노산 서열로 구성될 것으로 보이며 이배체는 69.018 kDa, 삼배체는 68.930 kDa 크기의 단백질을 만들 것으로 예상되었다. Vasa 재조합 단백질은 GST와 fusion되어 93 kDa 크기에서 관찰되었다. mRNA의 발현은 이배체에 비해 삼배체가 다소 높았다. Anti-Pw-VASA항체를 이용한 면역조직화학법을 수행한 결과 이배체와 삼배체는 다른 기관에서는 면역반응력을 보이지 않고 고환에서만 면역반응력을 볼 수 있었다. 결과를 종합해 보면 vasa 유전자는 이배체 삼배체 모두 가지고 있었으며 정상적으로 발현되었다.

Keywords

References

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