通过聚合酶链式反应（PCR）和限制性内切酶分析对小鼠和大鼠裸突变进行基因分型。

Genetic typing of the mouse and rat nude mutations by PCR and restriction enzyme analysis.

作者信息

Hirasawa T, Yamashita H, Makino S

机构信息

Center for Experimental Animals Development, Shionogi & Co., Ltd., Shiga, Japan.

出版信息

Exp Anim. 1998 Jan;47(1):63-7. doi: 10.1538/expanim.47.63.

DOI:10.1538/expanim.47.63

PMID:9498115

Abstract

A genetic typing method for the mouse and rat nude mutations by PCR and restriction fragment length polymorphism (RFLP) analysis was developed. Since restriction sites useful for RFLP analysis do not exist in the mouse nu and rat rnu mutations, artificial restriction sites were introduced by PCR with modified primers. Three genotypes in the mouse (nu/nu, nu/+ and +/+) or rat (rnu/rnu, rnu/+ and +/+) are rapidly differentiated with the PCR-RFLP assay. In addition, congenic nude strains can be efficiently established by using this assay. Finally, genetic mapping of the rnu locus was performed with microsatellite markers. The locus order on rat chromosome 10 was D10Mgh14-(2.0cM)-D10Mit2-(1.4cM)-rnu-(0.7cM++ +)-D10Mgh6-(2.7cM)-D10Mit8.

摘要

开发了一种通过聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）分析对小鼠和大鼠裸突变进行基因分型的方法。由于在小鼠nu和大鼠rnu突变中不存在可用于RFLP分析的限制性酶切位点，因此通过使用修饰引物的PCR引入人工限制性酶切位点。利用PCR-RFLP检测可快速区分小鼠（nu/nu、nu/+和+/+）或大鼠（rnu/rnu、rnu/+和+/+）的三种基因型。此外，使用该检测方法可有效建立同源裸鼠品系。最后，用微卫星标记对rnu基因座进行了遗传定位。大鼠10号染色体上的基因座顺序为D10Mgh14-（2.0厘摩）-D10Mit2-（1.4厘摩）-rnu-（0.7厘摩++ +）-D10Mgh6-（2.7厘摩）-D10Mit8。