睾丸注射法制备携带山羊<em>H-FABP</em>基因的转基因小鼠

阴彦辉，孙敏，陈庭锋，张亚妮，朱才业，李伟，李碧春

doi:10.3724/SP.J.1005.2012.00727

遗传 >

2012 , Vol. 34 >Issue 6: 727 - 735

DOI: https://doi.org/10.3724/SP.J.1005.2012.00727

研究报告

睾丸注射法制备携带山羊H-FABP基因的转基因小鼠

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扬州大学动物科学与技术学院, 江苏省动物遗传繁育与分子设计重点实验室, 扬州 225009

收稿日期: 2011-12-28

修回日期: 2012-02-17

网络出版日期: 2012-06-25

基金资助

国家转基因重大专项(编号：2008ZX08008-003, 2009ZX08008-003B)资助

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Generation of goat H-FABP overexpression transgenic mice by testicular injection

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Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Tech-nology, Yangzhou University, Yangzhou 225009, China

Received date: 2011-12-28

Revised date: 2012-02-17

Online published: 2012-06-25

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摘要

为探究睾丸注射法制备转基因动物的可能性, 文章将携带有山羊心脏型脂肪酸结合蛋白(H-FABP)和绿色荧光蛋白标签的重组载体经脂质体包裹后随机打点注射小鼠睾丸。对实验小鼠进行睾丸切片、精子荧光检测以及精子DNA检测, 证实外源基因在亲代小鼠体内成功表达。睾丸注射后小鼠与正常母鼠交配产生的F₁代, 以及F₁代自交产生的F₂代在不同水平均可检测到外源基因的成功表达, 阳性率分别为4%和30.23%。研究结果说明睾丸注射是一种制备转基因动物行之有效的方法, 且外源基因可以稳定遗传。该方法的完善和成熟对于动物转基因以及动物性状改良和育种具有理论和实践意义。

关键词： 睾丸注射; 转基因小鼠; 心脏型脂肪酸结合蛋白

本文引用格式

阴彦辉，孙敏，陈庭锋，张亚妮，朱才业，李伟，李碧春 . 睾丸注射法制备携带山羊H-FABP基因的转基因小鼠[J]. 遗传, 2012 , 34(6) : 727 -735 . DOI: 10.3724/SP.J.1005.2012.00727

Abstract

To explore the possibility of transgenic animals by testicular injection, the goat heart-type fatty acid binding protein (H-FABP) expression vector pEGFP-H-FABP was injected into the testis of 6 mice randomly by liposome mediated transfection. By detection of testis slice, sperm fluorescence and sperm DNA PCR, the exogenous gene was expressed in the parental mice. The exogenous gene was expressed at different levels in both the F₁ generation mice gave birthed by treated male mice and normal female mice and the F₂ generation mice generated by mating F₁ could be detected that the exogenous gene expressed at different levels with the positive rates of 4% and 30.23%, respectively. The results suggested that testicular injection, as an effective method to generate transgenic animal, could realize the stable integration of exogenous gene. The amelioration and maturity of testicular injection provides theoretical and practical significance in generation of transgenic animals and even in the animal trait improvement and breeding.

Key words： testicular injection; transgenic mice; heart-type fatty acid binding protein

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