转基因整合-自转基因罗非鱼 Hamilton（硬骨鱼纲：鲤科）的分析。

Transgene integration - an analysis in autotransgenic Labeo rohita Hamilton (Pisces: Cyprinidae).

机构信息

Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007, India,

出版信息

Fish Physiol Biochem. 2005 Apr;31(2-3):281-7. doi: 10.1007/s10695-006-0037-y.

DOI:10.1007/s10695-006-0037-y

PMID:20035471

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3233902/

Abstract

Transgenic Labeo rohita founder population was analyzed for the presence of autotransgene having histone 3 promoter and growth hormone (GH) cDNA (LRH3-GHcDNA) or total GH gene (LRH3-GH2.8) by PCR with transgene specific primers. Transgene specific amplification was seen with LRH3-GHcDNA in five out of seven individuals and all three fishes with LRH3-GH2.8, indicating their transgenic nature. Transgene integration was also studied by Southern hybridization of DNA isolated from blood of the transgenic fishes with two different probes (histone 3 promoter and cDNA of L. rohita). Autotransgene integration was confirmed in all PCR positive transgenic individuals. The site of integration of the transgene in the genome of the four transgenic fish could be determined by inverse PCR. Two individuals showed integration at the same site whereas in the remaining two individuals the integration sites were different.

摘要

采用转基因组特异性引物的 PCR 方法对携带组蛋白 3 启动子和生长激素 (GH) cDNA（LRH3-GHcDNA）或全长 GH 基因（LRH3-GH2.8）的转基因罗非鱼建立者群体进行了分析。在 7 个个体中的 5 个个体和 3 个具有 LRH3-GH2.8 的个体中均可见 LRH3-GHcDNA 的转基因特异性扩增，表明其为转基因个体。通过Southern 杂交从转基因鱼的血液中提取的 DNA 与两个不同的探针（组蛋白 3 启动子和罗非鱼 cDNA）也研究了转基因的整合。在所有 PCR 阳性的转基因个体中均证实了自体基因的整合。通过反向 PCR 可以确定 4 条转基因鱼基因组中转基因的整合位点。有两个个体显示出相同的整合位点，而另外两个个体的整合位点不同。