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半合子和纯合子转基因罗非鱼(尼罗罗非鱼)中与拷贝数相关的转基因表达和嵌合体细胞表达

Copy number related transgene expression and mosaic somatic expression in hemizygous and homozygous transgenic tilapia (Oreochromis niloticus).

作者信息

Rahman M A, Hwang G L, Razak S A, Sohm F, Maclean N

机构信息

Division of Cell Sciences, School of Biological Sciences, University of Southampton, United Kingdom.

出版信息

Transgenic Res. 2000 Dec;9(6):417-27. doi: 10.1023/a:1026517212807.

Abstract

Three lines of transgenic tilapia (Oreochromis niloticus) fish were generated with a construct containing a lacZ reporter gene spliced to a 4.7 kb 5' regulatory region of a carp beta actin gene. All these three lines contain different copy numbers of transgenes and the levels of lacZ expression were found to be related to transgene copy number. Mosaic patterns of somatic lacZ expression were observed in these three lines which differed between lines but were consistent within a line. We also observed that expression of the reporter gene in homozygous transgenic fish was approximately two-fold greater than in the hemizygous transgenics. Analysis of expression of the reporter gene on a tissue-to-tissue basis demonstrated that lacZ expression of the reporter gene in stably transformed fish occured with variable intensity in different organs and tissues and was also sometimes variable in different cells of the same tissue in Gland G2 generations of the transgenic lines.

摘要

利用一个构建体培育出了三代转基因罗非鱼(尼罗罗非鱼),该构建体包含一个与鲤鱼β-肌动蛋白基因4.7 kb 5'调控区域拼接的lacZ报告基因。这三代转基因鱼都含有不同拷贝数的转基因,并且发现lacZ表达水平与转基因拷贝数有关。在这三代转基因鱼中观察到了体细胞lacZ表达的镶嵌模式,三代之间有所不同,但同一代内是一致的。我们还观察到,纯合转基因鱼中报告基因的表达比半合子转基因鱼大约高两倍。基于组织对报告基因表达进行分析表明,在转基因系的G2代中,稳定转化鱼中报告基因的lacZ表达在不同器官和组织中的强度不同,并且在同一组织的不同细胞中有时也会有所变化。

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