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目的化学遗传学方法大规模筛选并初步鉴定所获得具有不同红系造血缺陷表型的斑马鱼突变体。方法乙基亚硝基脲(ENU)诱导雄性斑马鱼突变(founder),将其与野生型AB雌性斑马鱼交配产生F1代,源自不同来源的founder的F1代杂交产生F2家族。在F2代同家族内自交所产生的F3代胚胎中,用以βe1为探针,实施整体原位杂交实验,进行红系造血缺陷突变体筛选,并针对所筛选到的突变体在不同造血过程缺陷表型进行分类研究。结果和结论筛选得到4个βe1基因表达缺失突变体,其中2个为红系特异性造血缺陷突变体,另外2个突变体同时存在红系和淋系造血缺陷。
Objective To select and identify zebrafish mutants with different erythroid hematopoietic phenotype by large-scale screening and preliminary identification. Methods Ethyl nitrosourea (ENU) induced male zebrafish (founder) males to mate with wild-type AB female zebrafish to produce the F1 generation. F 1 generations of founder from different origins were crossed to produce the F2 family. F2 generation with the same generation of F3 generation embryos within the family, with βe1 as a probe, the implementation of the whole in situ hybridization screening of hematopoietic deficiencies of red screening, and screening for the mutant in different hematopoietic Process defect phenotypes were classified. RESULTS AND CONCLUSIONS: Four βe1 gene deletion mutants were screened, of which two were erythroid specific hematopoietic deficient mutants, while the other two mutants had both erythroid and lymphoid haematopoietic deficiencies.