陈霞 仝欣 潘筱云 许译丹 李建民.高表达人源SP15 对小鼠生长和生殖的影响[J].,2014,14(21):4009-4013 |
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 andReproduction of Mice |
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DOI: |
中文关键词: 胞苷脱氨酶 NYD-SP15 转基因小鼠 生长 生殖 |
英文关键词: Cytidine deaminase NYD-SP15 Transgenic mice Growth Reproduction |
基金项目:国家自然科学基金项目(31171443) |
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中文摘要: |
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。 |
英文摘要: |
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. |
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