TitleC6orf120基因敲除大鼠的制备及基因型鉴定
Other TitlesPreparation and genotype identification of C6orf120 gene knockout rat
Authors张健
张曼卡
马慧敏
宋心成
何玲玲
李鑫
Affiliation北京大学地坛医院教学医院中西医结合中心
首都医科大学附属北京地坛医院中西医结合中心
首都医科大学附属北京地坛医院传染病研究所
KeywordsC6orf120
基因敲除
大鼠
C6orf120
Gene knockout
Rat
Issue Date2019
Publisher胃肠病学和肝病学杂志
Abstract目的建立稳定的C6orf120基因敲除SD大鼠模型,观察基因敲除对大鼠生长发育的影响,探索该基因的功能及其在肝脏损伤中的作用。方法采用TALEN基因敲除技术敲除基因C6orf120,获得杂合子(C6orf120~(+/-))大鼠,将C6orf120~(+/-)大鼠进行杂交后获得纯合子(C6orf120~(-/-))大鼠。采用DNA Analyzer 3730XL测序技术对大鼠的基因型进行鉴定。观察子代大鼠的表型变化及对大鼠子代生育情况进行统计。随机处死8周龄C6orf120~(-/-)大鼠和野生型(wild type,WT)大鼠,留取大鼠的心、肝、脾、肺、肾、胸腺、淋巴组织,通过Western blotting技术检测C6orf120蛋白在大鼠各组织的表达情况。结果获得了稳定遗传的C6orf120~(-/-)基因型大鼠。Western blotting技术证明基因C6orf120在C6orf120~(-/-)大鼠体内全部删除,而C6orf120蛋白主要在WT大鼠的肝组织中高表达,在脾脏高表达,在肺、胸腺、淋巴组织中低表达。观察发现,WT大鼠和C6orf120~(-/-)大鼠的外观和行为表现上差异无统计学意义。对子代大鼠性别进行观察发现,C6orf120~(-/-)大鼠的雌性数量明显多于雄性。结论 C6orf120基因敲除大鼠模型成功构建,该模型的构建为探索该基因在肝损伤中作用奠定了基础。C6orf120基因可能参与大鼠的生长发育过程。
Objective To establish a stable C6orf120 gene knockout rats model, to explore the functions of C6orf120 gene in liver injury and to observe the effects of C6orf120 gene on growth and development. Methods C6orf120~(+/-) rats were made by the technology of TALEN gene knockout. The C6orf120~(-/-) rats were successfully obtained after the hybridization of C6orf120~(+/-) genotype rats. The rat genotype was identified by DNA Analyzer 3730 XL sequencing technology. The phenotypic changes and reproductive status of offspring rats were detected. Eight weeks old male wild type(WT) and C6orf120~(-/-) rats were selected and sacrificed randomly. Heart, liver, spleen, lung, renal, thymus and lymph were collected to analyze the expression of C6orf120 gene at indicated tissues by Western blotting. Results C6orf120~(-/-) rats were produced. The Western blotting results showed that C6orf120 gene was knockout absolutely in C6orf120~(-/-) rats. C6orf120 protein was mainly expressed in liver and spleen, and weakly expressed in lung, thymus and lymph. There was no remarkable difference on the appearance and behavior of all genotype rats. By observing the sex of the offspring rats, the number of females in C6orf120~(-/-) rats was more than males. Conclusion The rat model of C6orf120 knockout is successfully constructed, and the construction of this model laid a foundation for exploring the role of this gene in liver injury. C6orf120 gene may be involved in growth and development of rats.
URIhttp://hdl.handle.net/20.500.11897/562582
ISSN1006-5709
Appears in Collections:北京地坛医院 

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