A NOVEL MULTISPECIFIC COMPETITOR FRAGMENT FOR QUANTITATIVE PCR ANALYSIS OF CYTOKINE GENE-EXPRESSION IN RATS

被引：166

作者：

SIEGLING, A

LEHMANN, M

PLATZER, C

EMMRICH, F

VOLK, HD

机构：

[1] HUMBOLDT UNIV BERLIN,MED FAC CHARITE,INST MED IMMUNOL,D-10098 BERLIN,GERMANY

[2] MAX PLANCK SOC,RHEUMATOL IMMUNOL RES UNIT,D-91054 ERLANGEN,GERMANY

[3] UNIV ROSTOCK,INST MED BIOCHEM,ROSTOCK,GERMANY

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1994年 / 177卷 / 1-2期

关键词：

CYTOKINE; RAT; POLYMERASE CHAIN REACTION; GENE EXPRESSION; COMPETITOR CONTROL FRAGMENT;

D O I：

10.1016/0022-1759(94)90139-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Competitive polymerase chain reaction (PCR) is a sensitive method for quantification of cytokine mRNA expression. Co-amplification of an internal standard serves as control for comparing the efficiency of PCR in different samples. We have developed a novel control fragment for multiple analyses of rat cytokine gene expression containing primers for IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, TNF-alpha, TGF-beta(1), IFN-gamma and MIP-2. Additional primers were incorporated to analyse the content of T cells (CD3), activated T cells (CD25) and housekeeping genes (beta-actin and HPRT). As an example we demonstrate analysis of IL-2 mRNA expression in small pieces of kidney tissue obtained from rats after kidney allotransplantation. The IL-2 expression decreased tenfold during treatment with an anti-rat CD4 monoclonal antibody as compared to untreated animals.

引用

页码：23 / 28

页数：6

共 12 条

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