| Caveolin-1基因敲除小鼠子代基因型的鉴定及繁育方法 |
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| 引用本文： | 周胜强,罗东,黄素芬,易健,刘柏炎.Caveolin-1基因敲除小鼠子代基因型的鉴定及繁育方法[J].实验动物与比较医学,2016,24(3):228-232. |
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| 作者姓名： | 周胜强 罗东 黄素芬 易健 刘柏炎 |
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| 作者单位： | 湖南中医药大学, 长沙 410208;湖南中医药大学, 长沙 410208;湖南中医药大学, 长沙 410208;湖南中医药大学第一附属医院, 长沙 410007;湖南中医药大学, 长沙 410208 |
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| 基金项目： | 国家自然科学基金(NO.81273989);湖南省教育厅创新平台项目(NO.12K089) |
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| 摘 要： | 目的 探讨小凹蛋白-1(caveolin-1)基因敲除小鼠的鉴定方法与最优繁育方式,为深入研究caveolin-1在脑缺血损伤修复中的作用提供理想的动物模型。方法 将引进的caveolin-1基因敲除小鼠饲养于SPF级实验室,煮沸裂解法提取鼠尾组织基因组DNA,根据美国杰克逊实验室(Jackson Laboratory)提供的引物序列进行PCR反应检测其基因型,采用caveolin-1+/-杂合子互交、杂合子与caveolin-1-/-纯合子杂交(正交及反交)、纯合子互交4种不同的交配方式,观察亲代小鼠的受孕率、子代小鼠的外形特征及纯合率。结果 琼脂糖凝胶电泳显示PCR产物分子量大小约200 bp和661 bp,与预期的目的基因片段分子量大小一致,成功鉴定了caveolin-1基因敲除小鼠的不同基因型;不同交配方式的繁殖结果基本符合孟德尔遗传规律,且雌性、雄性caveolin-1-/-纯合鼠具有一定的繁殖能力,三种不同基因型小鼠的外形特征无明显差异。结论 煮沸裂解法提取基因组DNA、PCR法能够快速可靠鉴定caveolin-1基因敲除小鼠的基因型;caveolin-1杂合子小鼠互交与纯合子互交相结合的繁育方法可能是短期内获得足量纯合子子鼠与同源野生子鼠的较好方式。
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| 关 键 词： | 小凹蛋白-1 基因敲除 小鼠 鉴定 繁育方法 |
| 收稿时间： | 2015/12/11 0:00:00 |
Genotype identification and breeding method of caveolin-1 gene knockout mice |
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| ZHOU Sheng-qiang,LUO Dong,HUANG Su-feng,YI Jian and LIU Bai-yan.Genotype identification and breeding method of caveolin-1 gene knockout mice[J].Laboratory Animal and Comparative Medicine,2016,24(3):228-232. |
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| Authors: | ZHOU Sheng-qiang LUO Dong HUANG Su-feng YI Jian and LIU Bai-yan |
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| Affiliation: | Hunan University of Chinese Medicine, Changsha 410208, China;Hunan University of Chinese Medicine, Changsha 410208, China;Hunan University of Chinese Medicine, Changsha 410208, China;The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007;Hunan University of Chinese Medicine, Changsha 410208, China |
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| Abstract: | Objective To investigate the identification and optimal breeding method of caveolin-1 knockout mice, and provide an ideal animal model for further study of the role of caveolin-1 in cerebral ischemic injury and repair. Methods The introduced caveolin-1 gene knockout mice were reared in the SPF laboratory and genomic DNA was extracted from mouse tail tissue by the method of boiling lysis. According to the primer sequences provided by the Jackson Laboratory of America for polymerase chain reaction (PCR) to detect the genotypes, with the four different ways of mating:caveolin-1+/-heterozygote intercrossing, heterozygous and homozygous caveolin-1-/- hybrid (orthogonal and pay) as well as homozygous intercrossing. The pregnancy rate, shape characteristics of the filial generation mice and homozygous rate of the parental mice were observed. Results Agarose gel electrophoresis results indicated that the size of molecular weight of the PCR products was about 200 bp and 661 bp, which were consistent with the expected target gene fragment, and identified caveolin-1 gene knockout mice of different genotypes successfully. The results of different mating patterns are basically in agreement with Mendel rule, and the female and male aveolin-1-/- homozygous mice had a certain ability to reproduce, three different genotypes of mice had no significant differences between the shape features. Conclusions PCR can fast and reliably identify the genotypes of caveolin-1 knockout mice using genomic DNA through the method of boiling lysis. Combining the breeding methods of intercrossing of caveolin-1 heterozygous mice and intercrossing of caveolin-1 homozygous mice may be a good way to obtain enough homozygous mice and homologous wild type mice in a short period. |
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| Keywords: | Caveolin-1 Gene knockout Mice Identification Breeding methods |
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