高表达人源SP15对小鼠生长和生殖的影响

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。

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

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 systemand caused the lower birth rate of positive F2 offspring than F1.