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一种简单快速的方法来确定uPA转基因SCID小鼠的纯合性。

A simple and rapid method to determine the zygosity of uPA-transgenic SCID mice.

作者信息

Meuleman P, Vanlandschoot P, Leroux-Roels G

机构信息

Center for Vaccinology, Ghent University, De Pintelaan 185, Ghent 9000, Belgium.

出版信息

Biochem Biophys Res Commun. 2003 Aug 22;308(2):375-8. doi: 10.1016/s0006-291x(03)01388-3.

Abstract

Successful transplantation of xenogeneic hepatocytes into uPA-transgenic SCID mice depends on the zygosity of the recipient mice. Normally, the difference between homozygous and heterozygous animals is determined via a quantitative Southern blot. We sequenced a part of the mouse genome that is eliminated upon integration of the transgene in the genome. Based on that sequence we developed a multiplex PCR that allows the unambiguous discrimination of negative, heterozygous, and homozygous uPA-transgenic SCID mice in a single day procedure. The speed of the procedure is an essential quality because transplantation of xenogeneic hepatocytes into uPA-SCID mice should be done as soon as possible after birth.

摘要

将异种肝细胞成功移植到uPA转基因SCID小鼠体内取决于受体小鼠的纯合性。通常,纯合子和杂合子动物之间的差异通过定量Southern印迹法来确定。我们对转基因整合到基因组时被消除的小鼠基因组的一部分进行了测序。基于该序列,我们开发了一种多重PCR方法,可在一天内明确区分阴性、杂合子和纯合子uPA转基因SCID小鼠。该方法的速度是一项重要特性,因为将异种肝细胞移植到uPA-SCID小鼠体内应在出生后尽快进行。

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