grx-1基因敲除鼠的繁殖及子代基因型鉴定.doc

Grx-1基因敲除鼠的繁殖及子代基因型鉴定 刘秋彤，韩文莉，郭春宝，邓 春 (重庆医科大学附属儿童医院，重庆400014） [摘要] 目的 探讨glutaredoxin-1(Grx-1)基因敲除小鼠的优化繁殖及子代鼠的鉴定方法，为进一步研究Grx-1在支气管肺发育有不良(BPD)中的作用奠定基础。方法 将从美国哈佛医学院引进的纯合子Grx-1基因敲除小鼠进行饲养繁殖，繁殖出的子代中将出现纯合子、杂合子以及野生型3种基因型。从出生起观察其生长发育情况，2周龄时从子鼠尾部提取基因组DNA，用PCR方法扩增目的基因片段，琼脂糖凝胶电泳结果判定基因型。结果 Grx-1纯合子小鼠的饲养繁殖取得成功，获得了一批Grx-1基因敲除纯合子小鼠。结论 正确的饲养繁殖以及鉴定方法是获得Grx-1基因敲除纯合子小鼠的有效途径，为相关研究提供动物实验模型奠定了基础。 [关键词] 谷氧还蛋白；基因敲除；基因型鉴定；氧化应激 [中图分类号] R-332；Q95-33 Breeding and identification of glutaredoxin-1 knockout mice Liu Qiutong, Han Wenli, Guo Chunbao, Deng Chun (Children’s Hospital Affiliated to Chongqing Medical University,Chongqing 400014, China) [Abstract] Objective To explore the optimal way of breeding and identification of glutaredoxin-1（Grx-1） knockout mice and to establish an animal experimental model for further study on the important role of Grx-1 gene. Methods Breeding homozygote genotype mice obtained from Harvard Medical School, reproductive success of their offspring will appear in three genotypes: wild-type, heterozygous and homozygous. To observe the offspring’s growth and development, genomic DNA of each mouse was extracted from the tail of two-week-old mice. PCR method was employed to amplify the Grx-1 gene fragment, and the gene type was observed by electrophoresis. Results Breeding and reproducing were successful and gained number of homozygous knockout mice. Conclusion Appropriate methods of breeding and identification is an effective way to obtained homozygous Grx-1 knockout mice wich could provide as an experimental model of relevant research. [Key words] Glutaredoxin; gene knockout; gene identification; oxidative stress. Corresponding author: Deng chun, Tel: (023 E-mail:dengcgcb@ 谷氧还蛋白(Grx)是体内重要的氧化还原物质之一，在细胞内氧化还原平衡的调控以及抵抗氧化应激损伤过程中发挥着重要作用[1]。人类有两种谷氧还蛋白同工酶(Grx-l和Grx-2)[2]，Grx-l定位于胞液中，Grx-2分布在线粒体和核膜上。Grx-l的抗氧化作用明显强于Grx-2[3]，因此对Grx-1氧化还原的研究有助于明确氧化应激相关疾病的致病机理。近来也有文献报道Grx-2在脊椎动物大脑的发育方面至关重要[4]，总之Grx家族是人类生理和病理过程中至关重要的氧化还原物质，但其在各种疾病中的具体机制仍不明确。研究发现导致支气管肺发育不良(BPD)的