高表达人源SP15对小鼠生长和生殖的影响

The Effects of Overexpressing Homo SP15 on Growth and Reproduction of Mice

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 observe the growth and reproduction in mice overexpressing homo SP15 gene and provide a transgenic animal model for the study of the biological function of SP15. Methods： NYD-SP15 transgenic vectors were constructed by inserting homo NYD-SP15 cDNA and the Cre fragment to the downstream of PCAG promoter. Microinjection was applied to get the mice expressing NYD-SP15 protein systemically. The positive offspring, which could stably express protein SP15, were obtained by mating NYD-SP15 transgenic mice with C57/BL6 mice and identified by PCR. Then the weights of positive transgenic mice were compared with their wildtype controls and the birth number of positive offspring was counted. Results： The NYD-SP15 transgenic overexpression vector was successfully constructed by PCR and plasmid sequence analysis. The NYD-SP15 transgenic mice were identified by PCR. It was indicated that overexpression of homo NYD-SP15 could not affect the body weight of mouse, since there was no significant difference between the weights of transgenic mice and littermate wild-type mice. What＇s more, the birth rate of positive F2 offspring was significantly lower than that of F1 in transgenic mice. Conclusion： Transgenic mice could express Homo NYD-SP15 normally and play biological function successfully. Since NYD-SP15 could be expressed in the testis, it was suggested that NYD-SP15 might influence the reproductive system and caused the lower birth rate of positive F2 offspring than F2.