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目的繁殖并鉴定HO-1基因敲除(HO-1-/-)小鼠。方法将引进的HO-1基因敲除杂合子小鼠,进行SPF级饲养,与C57BL/6野生型小鼠配种繁殖,提取子代小鼠的基因组DNA,PCR法特异性扩增HO-1基因片段,使用双脱氧测序法测得基因序列。子代小鼠出现3种基因型:野生型、杂合子型及纯合子型。杂合子小鼠进一步配种繁殖,以获得更多的基因敲除纯合子小鼠,纯合子小鼠用以课题研究。结果 HO-1基因敲除杂合子小鼠的饲养和繁殖均获得成功,获得了HO-1基因敲除纯合子和杂合子小鼠。结论正确的饲养繁殖及鉴定方法是从HO-1基因敲除杂合子小鼠中获得纯合子小鼠的有效途径。
Objective To breed and identify HO-1 knockout (HO-1 - / -) mice. Methods HO-1 gene knockout heterozygous mice were introduced and cultivated at SPF level to breed and breed with C57BL / 6 wild-type mice. The genomic DNA of offspring mice was extracted, and the HO-1 gene was amplified by PCR Fragments were determined using dideoxy sequencing. Three offspring mice showed three genotypes: wild-type, heterozygous and homozygous. Heterozygous mice further breeding breeding to get more gene knockout homozygous mice, homozygous mice for research. Results HO-1 gene knockout heterozygous mice were successfully fed and reproduced, and HO-1 knockout homozygous and heterozygous mice were obtained. Conclusion The correct method of breeding, breeding and identification is an effective way to obtain homozygous mice from HO-1 knockout heterozygous mice.