Apr3条件性基因敲除小鼠模型的建立及初步表型研究

发布时间：2018-01-08 21:23

本文关键词：Apr3条件性基因敲除小鼠模型的建立及初步表型研究　出处：《上海交通大学学报(医学版)》2015年08期 　论文类型：期刊论文

【摘要】：目的建立凋亡相关蛋白3(Apr3)条件性基因敲除小鼠模型,并初步研究其表型。方法设计基因敲除方案,获得Apr3flox(+/-)嵌合体小鼠,繁殖获得Apr3(-/+)小鼠后,再自交得到Apr3基因全身敲除Apr3(-/-)纯合小鼠和对照野生型小鼠。提取小鼠鼠尾组织基因组DNA,PCR扩增野生型和敲除基因片段,在DNA水平判断小鼠的基因型;提取小鼠肝脏RNA和蛋白,利用RT-PCR和Western blotting技术检验Apr3基因的表达;观察基因敲除小鼠的遗传性状、体质量变化趋势以及可能影响生长发育和代谢的指标如血糖、血脂、肝功能;对小鼠溶酶体富集的主要脏器行苏木精-伊红染色(H-E染色)观察。结果成功建立Apr3基因敲除小鼠模型,其中Apr3(-/-)小鼠存活并且可育,在2月龄时体质量落后于野生型小鼠。肝功能检测结果显示,2月龄Apr3(-/-)小鼠谷草转氨酶(AST)较野生型小鼠升高(P0.05);H-E染色结果显示,肝组织出现明显肝细胞水肿样变性和汇管区炎症细胞浸润。结论初步建立了Apr3条件性基因敲除小鼠模型;Apr3基因敲除引起2月龄小鼠体质量落后、肝脏组织形态学异常和AST升高。
[Abstract]:Objective to establish the apoptosis related protein 3 (Apr3) gene conditional knockout mice, and to study the method of gene knockout phenotype. The design scheme, Apr3flox (+ / -) chimeric mice, Apr3 (- / +) mice reproduction, and self Apr3 gene knockout body Apr3 (- / -) homozygous mice and controls the wild type mice. Genomic DNA was extracted from the mouse tail tissue, PCR amplification of wild-type and knockout gene fragment, determine the genotype of mice at DNA level; extraction of mouse liver RNA and protein expression by RT-PCR and Western blotting test of Apr3 gene; gene knockout mice to observe genetic traits, body weight changes and trends may affect the growth and metabolism indexes such as blood glucose, blood lipid, liver function; the main organs of mouse lysosomal accumulation of hematoxylin eosin staining (H-E staining) were observed. Results the successful establishment of Apr3 gene knockout mice, The Apr3 (- / -) mice survival and fertility, in February at the age of mass behind the wild type mice. Liver function test results showed that the February age Apr3 (- / -) mice aspartate aminotransferase (AST) increased compared with wild-type mice (P0.05); H-E staining showed that the liver tissue appeared obvious liver cell edema like periportal degeneration and inflammatory cell infiltration. Conclusion the initial establishment of the Apr3 conditional gene knockout mice; Apr3 gene knockout mice caused by body mass in February behind, liver tissue morphological abnormalities and AST increased.

【作者单位】：上海交通大学医学院附属新华医院上海市儿科医学研究所内分泌遗传代谢病研究室;
【基金】：国家自然科学基金(81071121)~~
【分类号】：R-332;R363
【正文快照】： 凋亡相关蛋白3(apoptosis related protein 3,APR3)为功能尚未明确的溶酶体膜蛋白(lysosomalmembrane proteins,LAMPs)。目前,对该基因功能的研究较少。最初对CD34+造血干细胞中未定义表达基因的研究表明,APR3参与造血干细胞的形成和分化[1]。有研究在全反式维甲酸处理的人早

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