CAJ | 학술논문

目的斑马鱼cloche172突变体的基因鉴定.方法采用化学诱变剂N-乙基-N-亚硝基脲(N-ethyl-N-nitrosourea,ENU)诱变雄性野生型AB斑马鱼,实施大规模正向遗传学方法筛选髓系细胞标记物lysozyme C(lyC)表达缺失突变体,并对其中一个名为cloche~(172)突变体进行大体形态学观察,基因定位克隆和遗传学互补实验.结果大规模正向遗传学筛选出4个lyc表达缺失突变体,其中一个cloche~(172)突变体受精后3 d(day post fertilization,dpf)时期大体形态改变与已知但基因不明的cloche突变体相似.遗传学互补实验观察到有1/4胚胎出现类似cloche突变体的表型.同时,cloche~(172)突变体基因定位在13号染色体近端粒的区域,与cloche突变体基因定位区域一致.而o-dianisdine实验证实cloche~(172)突变体有较多红细胞聚集,cloche突变体仅有少量在尾部存在.结论 cloche~(172)基因与cloche基因定位一致,cloche~(172)突变体是一个新的点突变位点的cloche突变体.
목적 반마어cloche172돌변체적기인감정.방법 채용화학유변제N-을기-N-아초기뇨(N-ethyl-N-nitrosourea,ENU)유변웅성야생형AB반마어,실시대규모정향유전학방법 사선수계세포표기물lysozyme C(lyC)표체결실돌변체,병대기중일개명위cloche~(172)돌변체진행대체형태학관찰,기인정위극륭화유전학호보실험.결과 대규모정향유전학사선출4개lyc표체결실돌변체,기중일개cloche~(172)돌변체수정후3 d(day post fertilization,dpf)시기대체형태개변여이지단기인불명적cloche돌변체상사.유전학호보실험관찰도유1/4배태출현유사cloche돌변체적표형.동시,cloche~(172)돌변체기인정위재13호염색체근단립적구역,여cloche돌변체기인정위구역일치.이o-dianisdine실험증실cloche~(172)돌변체유교다홍세포취집,cloche돌변체부유소량재미부존재.결론 cloche~(172)기인여cloche기인정위일치,cloche~(172)돌변체시일개신적점돌변위점적cloche돌변체.
Objective To perfonn the genetic idencification of cloche~(172) mutant zebrafish. Methods The chemical mutager.N-ethyl-N-nitrosourea (ENU) was used to treat the AB stain male fish. Large-scale forward genetic screening was carried out to search for lyC-deficient zebrafish mutant by WISH. The morphology changes of the embryos at 3 days postfertilization (3dpf) stage were observed and the cloche~(172) gene was identified by mapping and complementation test. Results We selected 4 lyC-deficient zebrafish by WISH. cloche~(172) mutant showed morphological changes similar to cloche mutant in 3dpf stage. One fourth of the embryos showed cloche phenotype as found in complementation test, and the cloche~(172) gene was mapped on the telomere of zebrafish 13 chromosome where cloche gene was located. Numerous red blood cells were observed in the cloche~(172) mutant, wlule only a few cells were found in the cloche mutant in the tail region by o-dianisdine staining. Conclusion cloche~(172) gene which is responsible for the phenotype of cloche mutant may be a novel point mutation allele of the cloche mutant.