SETD4基因敲除小鼠的构建及鉴定

doi:10.13418/j.issn.1001-165x.2016.02.017

中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (2): 202-207.doi: 10.13418/j.issn.1001-165x.2016.02.017

SETD4基因敲除小鼠的构建及鉴定

黄穗，　黄梦怡，　钟玙沄，　雷烨铭，　赵舒祺，　蔡军伟，　姜勇，　刘靖华

广东省蛋白质组学重点实验室，南方医科大学病理生理学教研室，广州 510515

收稿日期:2015-12-25 出版日期:2016-03-25 发布日期:2016-04-14
通讯作者: 刘靖华，教授，博士生导师，Tel：（020）61648172-810，E-mail:liujhua@fimmu.com；姜勇，教授，博士生导师，Tel：020-61648231，E-mail: jiang48231@163.com
作者简介:黄穗（1991-），女，广东惠州人，在读硕士，主要从事脓毒症休克机制方向的研究，Tel:（020）61648172-136，E-mail：13430259775@163.com
基金资助:
国家自然科学基金项目（No. 81072425，No.81471901）；广东省自然科学基金重点项目（ 2015A030311031）

Establishment and Identification of SETD4gene knockout mice

HUANG Sui, HUANG Meng-yi, ZHONG Yu-yun, LEI Ye-ming, ZHAO Shu-qi, CAI Jun-wei, JIANG Yong, LIU Jing-hua

Key laboratory for Functional Proteomics of Guangdong Province，Department of Pathophysiology，Southern Medical University，Guangzhou 510515

Received:2015-12-25 Online:2016-03-25 Published:2016-04-14

摘要/Abstract

摘要：

目的　构建并鉴定SETD4基因敲除小鼠，为研究SETD4的生物学功能提供动物模型。方法　将引进的SETD4flox/+小鼠与EIIa-Cre小鼠进行杂交繁殖，得到基因型为SETD4+/-.EIIa-Cre的小鼠；再与C57BL/6小鼠杂交去除Cre酶，获得杂合子SETD4+/-小鼠；该小鼠自交获得纯合子SETD4-/-小鼠。通过PCR法鉴定子代小鼠的基因型；RT-PCR、荧光定量PCR方法鉴定纯合子的SETD4基因敲除小鼠SETD4 mRNA表达情况；HE染色观察小鼠肝、肺组织的形态学变化。结果　PCR结果表明子代小鼠的基因型符合SETD4-/-；纯合子基因敲除小鼠SETD4 mRNA水平显著低于野生型小鼠；SETD4基因敲除小鼠肝、肺组织的形态学特征与野生型小鼠相比无明显差异。结论　本研究基于Cre/loxp系统，成功构建并鉴定了SETD4基因敲除小鼠。

关键词: SETD4, 基因敲除, Cre/loxp系统, KO-First技术

Abstract:

Objective　To study the function of SETD4， the SETD4 gene knockout homozygous mice has been established. 　Methods　SETD4flox/+ mice and EIIa-Cre mice were interbred，the offspring of which was genotyping SETD4+/-.EIIa-Cre were crossed with C57BL/6 mice to obtain the mice with the SETD4+/- genotype，SETD4+/- heterozygous mice were inbred and then the SETD4-/- homozygous mice were gained. PCR was used to identify the genotype of the offspring，the expression of SETD4 mRNA was detected by RT-PCR and qPCR，and morphological changes of liver and lung were observed by HE staining.　Result　PCR results showed genotypes of the offspring of SETD4 gene knockout mice was in accordance with SETD4-/-. Compared with the wild type mice，expression of SETD4 mRNA in SETD4 gene knockout homozygous mice was significantly decreased，and morphological characteristics of liver and lung in SETD4 gene knockout homozygous mice had no significant changes.　Conclusion　Wehave successfully generated SETD4 gene knockout homozygous mice which can be used for study ofSETD4 function.

Key words: SETD4, gene knockout, Cre/loxp, KO-First technology

黄穗，　黄梦怡，　钟玙沄，　雷烨铭，　赵舒祺，　蔡军伟，　姜勇，　刘靖华. SETD4基因敲除小鼠的构建及鉴定[J]. 中国临床解剖学杂志, 2016, 34(2): 202-207.

HUANG Sui, HUANG Meng-yi, ZHONG Yu-yun, LEI Ye-ming, ZHAO Shu-qi, CAI Jun-wei, JIANG Yong, LIU Jing-hua. Establishment and Identification of SETD4gene knockout mice[J]. Chinese Journal Of Clinical Anatomy, 2016, 34(2): 202-207.

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SETD4基因敲除小鼠的构建及鉴定

Establishment and Identification of SETD4gene knockout mice

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