miR-15a/16-1基因敲除小鼠的鉴定及其脾脏免疫细胞频率分析
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国家自然科学基金(81273214);江苏省普通高校研究生创新计划(CXLX13-924)


Identification of miR-15a/16-1-/- mice and analysis of their splenic cellular immune function
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    摘要:

    目的:探讨miR-15a/16-1基因敲除小鼠的繁育与子代鼠的鉴定,及其脾脏免疫细胞频率。方法:从子鼠鼠尾中提取基因组DNA,采用PCR方法进行基因型鉴定。miR-15a/16-1基因敲除小鼠眼眶取血,用动物血液分析仪进行全血细胞分析。采用流式细胞仪分析miR-15a/16-1基因敲除小鼠脾脏中免疫细胞频率。结果:miR-15a/16-1+/+-miR-15a/16-1+/--miR-15a/16-1-/- 各表型小鼠互交繁殖结果基本符合孟德尔遗传规律。流式结果显示,miR-15a/16-1基因敲除小鼠与野生型小鼠相比,CD19+-NKG2D+ 细胞增多,其余无明显差异。全血细胞分析结果显示淋巴细胞增多。结论:实验所用PCR方法可以鉴定miR-15a/16-1-/-小鼠;miR-15a/16-1-/-小鼠的获得为进一步探究miR-15a/16-1的作用提供了较理想的动物模型;敲除miR-15a/16-1基因,小鼠脾脏CD19+-NKG2D+细胞增多。

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    Objective:To breed and identify the offspring of positive miR-15a/16-1 gene knockout mice and analysis of their splenic cellular immune function. Methods:MiR-15a/16-1 knockout mice were paired in different ways. Genomic DNA was isolated from tails and analyzed by PCR. Complete blood analysis was performed in the blood which was collected from the orbit of mice. Analysis of cellular immune function in spleen was performed in miR-15a/16-1 gene knockout mice by FCM. Results:The positive rate with miR-15a/16-1 gene of filial generation mice met Mendelian genetic law. Compare with wild-type mice,CD19+ and NKG2D+ cells of spleen in miR-15a/16-1 gene knockout mice were all up-regulated expression detected by FCM,although there were not significant changes of frequencies of other surface markers. Complete blood analysis showed that lymphocytes increased. Conclusion:PCR methods can be used to identify the genotype of the miR-15a/16-1-/- mice,and to establish an animal experimental model to further study the important role of miR-15a/16-1. CD19+ and NKG2D+ cells of spleen in miR-15a/16-1 gene knockout mice were increased.

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沈娅婷,胡春霞,陈文艳,李晓敏,刘 浩,李国利,龚卫娟,贾筱琴. miR-15a/16-1基因敲除小鼠的鉴定及其脾脏免疫细胞频率分析[J].南京医科大学学报(自然科学版),2016,(3):330-334

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  • 收稿日期:2015-10-23
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  • 在线发布日期: 2016-03-24
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