用于RasH2转基因小鼠基因分型和纯合性检测的PCR方法。

PCR method for genotyping and zygosity-testing of RasH2 transgenic mice.

作者信息

Suemizu Hiroshi, Kito-Maruyama Chika, Sotomaru Yusuke, Ogura Tomoyuki, Hioki Kyoji, Ohnishi Yasuyuki, Tamaoki Norikazu

机构信息

Central Institute for Experimental Animals, Kanagawa, Japan.

出版信息

Exp Anim. 2004 Oct;53(5):463-6. doi: 10.1538/expanim.53.463.

DOI:10.1538/expanim.53.463

PMID:15516796

Abstract

In short-term carcinogenicity testing using CB6F1-TgrasH2 mice, sibling nonTgrasH2 mice are used as a negative control. However, selection of TgrasH2 and nonTgrasH2 mice has been performed by PCR with only transgene specific primers by the conventional method. Therefore, the conventional method involves the risk of false negative results due to reaction failure, and contamination with TgrasH2 mice in the control mice group. Based on the nucleotide sequence information around the pre-integration site, we developed a genotyping method for distinguishing not only TgrasH2 mice (hemizygous for the Tg allele) but also nonTgrasH2 (homozygous for the nonTg allele) in a positive manner.

摘要

在使用CB6F1-TgrasH2小鼠进行的短期致癌性测试中，同窝非TgrasH2小鼠用作阴性对照。然而，按照传统方法，仅使用转基因特异性引物通过聚合酶链反应（PCR）来选择TgrasH2和非TgrasH2小鼠。因此，传统方法存在因反应失败导致假阴性结果以及对照小鼠组被TgrasH2小鼠污染的风险。基于整合前位点周围的核苷酸序列信息，我们开发了一种基因分型方法，能够以阳性方式区分TgrasH2小鼠（Tg等位基因半合子）和非TgrasH2小鼠（非Tg等位基因纯合子）。