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目的:探究高表达人源SP15对小鼠生长和生殖的影响,为研究NYD-SP15在生物体中的功能提供动物模型。方法:将人源NYD-SP15 cDNA以及Cre序列插入PCAG启动子下游,构建NYD-SP15转基因表达载体,并通过原核注射,构建全身性表达NYD-SP15的转基因小鼠;将获得的NYD-SP15转基因小鼠与C57/BL6小鼠交配,PCR鉴定转基因小鼠是否有Cre插入,筛选出人源NYD-SP15表达的小鼠,统计转基因小鼠体重变化和后代阳性情况。结果:通过PCR鉴定及公司测序验证,我们成功构建了NYD-SP15转基因过表达载体。并通过PCR鉴定小鼠基因型,筛选出可以表达人NYD-SP15的转基因小鼠。比较转基因小鼠与同窝野生型小鼠体重,发现转基因小鼠体重与同窝野生型小鼠相比无明显区别,说明在体内过表达NYD-SP15对小鼠体重无明显影响。通过对后代阳性小鼠出生情况统计发现F2代阳性小鼠出生率较F1代明显降低。结论:人源NYD-SP15在小鼠体内能正常表达,对其生长无明显影响,但其后代阳性出生率呈逐代下降趋势,推测该原因可能与NYD-SP15在睾丸中表达有关。
OBJECTIVE: To investigate the effects of high expression of human SP15 on growth and reproduction in mice and to provide an animal model for studying the function of NYD-SP15 in the organism. METHODS: NYD-SP15 cDNA and Cre sequence were inserted into the downstream of PCAG promoter to construct NYD-SP15 transgene expression vector. The recombinant plasmid NYD-SP15 was constructed by pronucleus injection. Mice were crossed with C57 / BL6 mice to identify if there was Cre insertion in transgenic mice. The mice with NYD-SP15 expression were screened out, and the changes of body weight and the positive cases were analyzed. Results: NYD-SP15 transgenic overexpression vector was successfully constructed by PCR identification and company sequencing. The genotypes of the mice were identified by PCR and the transgenic mice that could express human NYD-SP15 were screened out. Comparison of body weight between transgenic mice and wild-type littermates showed no significant difference in body weight between wild-type mice and wild-type littermates, indicating that overexpression of NYD-SP15 had no significant effect on body weight in mice. According to the statistics of the offspring positive mice, the birth rate of F2 positive mice was significantly lower than F1 generation. CONCLUSION: Human NYD-SP15 is normally expressed in mice and has no significant effect on its growth. However, the positive rate of its offspring is decreasing from generation to generation. This may be related to the expression of NYD-SP15 in testis.