Molecular characterization of the commonly used human androgen receptor expression vector, pSG5-AR

被引:19
作者
Litvinov, IV
Chang, C
Isaacs, JT
机构
[1] Johns Hopkins Univ, Sch Med, Sidney Kimmel Comprehens Canc Ctr, Grad Training Program Cellular & Mol Med, Baltimore, MD 21218 USA
[2] Univ Rochester, Med Ctr, Dept Pathol, George Whipple Lab Canc Res, Rochester, NY 14627 USA
关键词
5 '-UTR; 3 '-UTR; pSG5-AR; androgen receptor; CAG repeat; GGC repeat;
D O I
10.1002/pros.20027
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
BACKGROUND. pSG5-AR is a commonly used androgen receptor (AR) expression vector. However, a detailed molecular characterization of this construct has not been published. In this study, we describe the molecular organization of the above plasmid and analyze the AR transcript coded by the above construct. METHODS. pSG5-AR was sequenced by the standard dideoxy sequencing method. The obtained sequence was analyzed by the DS Gene software. RESULTS. A molecular map of the pSG5-AR plasmid as well as a map of the AR insert is provided. The AR transcript from pSG5-AR is lacking significant portions of the 5'- and 3'-untranslated region (UTR) and contains shorter than average CAG and GGC repeats. CONCLUSIONS. The above characteristics of the pSG5-AR are significant and may result in different regulation pattern of expression of the expressed AR in comparison to the endogenous gene. Such a conclusion is based on the knowledge that post-transcriptional regulation plays an important role in controlling the levels of AR in a cell. Such regulation is controlled by the regulatory elements within the 5'- and 3'-UTRs of the AR transcript. In addition, this vector might be coding for an AR with a different transactivational capacity than the endogenous AR gene. The transactivational activity of AR was shown to be affected by the size of the trinucleotide-repeat-regions (CAG and GGC) within the gene. (C) 2004 Wiley-Liss, Inc.
引用
收藏
页码:319 / 324
页数:6
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