Streamlined approach to functional analysis of promoter-region polymorphisms

被引：11

作者：

Coleman, SL ^{[1
]}

Hoogen-doorn, B ^{[1
]}

Guy, C ^{[1
]}

Smith, SK ^{[1
]}

O'Donovan, MC ^{[1
]}

Buckland, PR ^{[1
]}

机构：

[1] Univ Wales Coll Cardiff, Coll Med, Dept Psychol Med, Cardiff CF14 4XN, S Glam, Wales

来源：

BIOTECHNIQUES | 2002年 / 33卷 / 02期

关键词：

Cell culture - Cloning - Enzymes - Genes;

D O I：

10.2144/02332ht01

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We have developed a rapid method for identifying functional promoter-region polymorphisms. Using a modified pGL3 luciferase expression T-vector, we can amplify by PCR, clone, identify allelic pairs of a polymorphic gene promoter region, and prepare plasmids for cell culture 10 promoters (20 allele pairs) per week per researcher. By utilizing 96-well plate technology and an internal control plasmid expressing secreted alkaline phosphatase, each of these allele pairs can be tested for relative promoter activity in each of three cell lines (HEK293t, TE671, and JEG3) with similar resources.

引用

页码：412 / +

页数：4

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