果蝇CG3295基因是哺乳动物基因RMND5在果蝇中的同源物,其功能未知.利用P转座子敲除技术进行果蝇CG3295基因的敲除研究,将果蝇14234系的P转座子与△2—3系中的转座酶基因组合到同一个体,使P转座子在体内敲掉目的基因.从后代挑选不含P转座子和转座酶基因的500头雄蝇分别与缺失了CG3295基因的测交系交配,从后代筛选获得20个CG3295基因敲除候选系.进一步利用Hand-GFP系使候选系的心脏表达绿色荧光蛋白,在荧光显微镜下根据候选系是否表现心脏突变表型鉴定敲除系,最后获得了12个表现心脏突变表型的CG3295基因敲除系.
The CG3295 gene is a Drosophila homolog of the mammalian gene RMND5. Its function is yet unknown. In order to study the fu lction of CG3295, P-element knockout method was used to generate the CG3295 knockout fly. P-element from the 14234 line and the transposase gene from the A2-3 line were grouped into a fly for the knowdown of the gene. In the next generation, 500 males without P-element and the transposase gene were respectively crossed with the testcross line without the CG3295 gene. Twenty knockout candidate lines of CG3295 were obtained from the offspring. The hearts of the candidate lines were labeled with heart-specific green protein by the Hand-GFP line. The knockout lines with heart phenotype were identified under fluorescence microscope, and twelve knock-out lines with heart phenotype were finally obtained.