Wip1过表达转基因小鼠模型的制备与鉴定

doi:10.16431/j.cnki.1671-7236.2016.12.002

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (12): 3094-3100.doi: 10.16431/j.cnki.1671-7236.2016.12.002

Wip1过表达转基因小鼠模型的制备与鉴定

高倩, 胡言青, 唐懿挺, 牟玉莲

中国农业科学院北京畜牧兽医研究所, 北京 100193

收稿日期:2016-06-03 出版日期:2016-12-20 发布日期:2016-12-22
通讯作者: 牟玉莲 E-mail:mouyulian@caas.cn
作者简介:高倩(1986-),女,山东淄博人,博士生,研究方向:动物细胞工程和基因工程,E-mail:gaoqian36985@163.com
基金资助:
国家自然科学基金（31572378）

Establishment and Identification of Wip1 Over-expressed Transgene Mice Model

GAO Qian, HU Yan-qing, TANG Yi-ting, MU Yu-lian

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2016-06-03 Online:2016-12-20 Published:2016-12-22

摘要/Abstract

摘要：

本研究旨在构建Wip1过表达转基因小鼠，并对Wip1过表达转基因小鼠进行RNA水平上的筛选，为研究动脉粥样硬化的发生机制提供一种动物模型。通过逆转录聚合酶链式反应（RT-PCR）获得小鼠组织的cDNA，并以其为模板进行PCR扩增，对扩增片段进行胶回收，然后进行扩增片段和空载体的双酶切，将酶切后胶回收得到的Wip1基因全长与载体pcDNA3.1（+）进行连接，获得Wip1-pcDNA3.1（+）过表达载体，最终得到Wip1过表达转基因原代小鼠，并利用Southern blotting方法和实时荧光定量PCR方法对小鼠进行鉴定筛选。试验获得Wip1过表达转基因原代小鼠30只，利用Southern blotting方法鉴定出阳性小鼠11只，阳性率为36.67%，实时荧光定量PCR筛选Wip1过表达转基因小鼠阳性率为32.84%。以上结果表明，试验成功构建了Wip1-pcDNA3.1（+）过表达载体，并且经过Wip1基因在小鼠组织DNA和RNA水平上的鉴定筛选，成功获得了Wip1过表达转基因小鼠。

关键词: Wip1; 过表达; 小鼠模型

Abstract:

This study was aimed to build Wip1 over-expressed transgene mice, which could provide an animal model for clarifying the pathogenetic mechanism of atherosclerosis, and the transgene mice were screened at RNA level. Mice tissues cDNA were obtained by reverse transcription polymerase chain reaction (RT-PCR), which were used for the next PCR amplification template. The amplified fragments were used for gel recovery.Then, the amplified fragment and empty vector were double enzyme digested.Full-length gene of Wip1,which was obtained by gel recovery, was connected to pcDNA3.1(+) vector, and the Wip1-pcDNA3.1(+) over-expressed vector was constructed. Finally, Wip1 over-expressed transgene mice were obtained. The transgene mice were identified and screened by Southern blotting and qPCR. 30 Wip1 over-expressed transgene mice were got in this experiment, and the positive mice identified were 11 by Southern blotting, the positive rate was 36.67%. Moreover,the positive rate was 32.84% through screening by qPCR. Therefore, the Wip1-pcDNA3.1(+) over-expressed vector was constructed successfully, and the Wip1 over-expressed transgene mice were obtained through identifying and screening at DNA and RNA level of Wip1 in mice tissues.

Key words: Wip1; over-expression; mouse model

中图分类号:

高倩, 胡言青, 唐懿挺, 牟玉莲. Wip1过表达转基因小鼠模型的制备与鉴定[J]. 《中国畜牧兽医》, 2016, 43(12): 3094-3100.

GAO Qian, HU Yan-qing, TANG Yi-ting, MU Yu-lian. Establishment and Identification of Wip1 Over-expressed Transgene Mice Model[J]. , 2016, 43(12): 3094-3100.

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Wip1过表达转基因小鼠模型的制备与鉴定

Establishment and Identification of Wip1 Over-expressed Transgene Mice Model

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