Journal of Sericultural and Entomological Science (한국잠사곤충학회지)

Korean Society of Sericultural Science (한국잠사학회)
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Fundamental Study for RAPD-PR Analysis in the Silkworm, Bombyx mori

누에 RAPD-PCR 분석을 위한 기초연구

  • 황재삼 (농촌진흥청 잠사곤충연구소) ;
  • 이진성 (농촌진흥청 잠사곤충연구소, 농촌진흥청 연구관리국, 농촌진흥청 잠사곤충연구소, 농촌진흥청 잠사곤충연구소, 성균관대학교 유전공학과)
  • Published : 1996.06.01
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Abstract

Reproducible the random amplified polymorphic DNAs(RAPDs) patterns were obtained in the two silkworm strains(J111, Galwon) by adjusting concentration optimized of Taq DNA polymerase(one unit), dNTP(200$\mu$M), MgCl2(1.5mM) and template DNA(30ng). In addition, anealing temperature ranging 35$^{\circ}C$ to 42$^{\circ}C$ by the adjusted condition was investigated and fixed at 35$^{\circ}C$ in this study. Variation among individuals and between male and female of Jam 113 strain was not authorized. DNA polymorhpisms among silkworms were authorized by five RAPD markers using OPM04 random primer. Using the primer showing polymorhpims between parents(J111, Galwon) in thirty three individuals, RAPD-PCR for F2 analysis was performed and segregated 3 : 1 in the F2 population. Consequently, RAPDs detected in the parents were obtained as genetic markers, which can be used for construction of genetic map for this industrially particular insect, silkworm Bombyx mori

누에 RAPD-PCR 최적조건에 대해 실험한 결과, 중합효소 연쇄반응(PCR)의 최적조건은 반응액 25$\mu$l에 대해 주형 DNA 30ng, dNTP mixture 200$\mu$M, primer 300mM, Taq DNA Polymerase 1.0unit 및 Mg2+ 1.5mM로 판단되었으며, 또 상기의 실험조건을 기본으로 하여 annealing 온도를 탐색한 결과 35$^{\circ}C$-42$^{\circ}C$에서 DNA의 증폭이 안정적임이 밝혀졌다. 또한 동일한 품종내에서 개체간 및 암수간의 DNA 다형성은 인정되지 않았고, 품종간 DNA 다형성은 OPM 04 primer를 사용한 결과 5개의 RAPD 마커로 인정 되었다. 양친간 다형성이 인정된 primer를 사용하여 F2 33개체에 대해 DNA를 증폭시킨 결과 800 bp band가 3 :1 로 분리되었으므로 누에 품종간 유전적 유연관계 및 유전자 지도작성의 가능성을 제시했다.

Keywords

References

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