A competitive reverse transcription–PCR to study apolipoprotein ε gene expression

Abstract We developed a rapid and simple competitive reverse transcription-polymerase chain reaction for the quantification of apoε mRNA in human monocyte-derived macrophages. The method was applied, and its reliability was shown in patients with the familial lipoprotein disorder, type III hyperlipoproteinemia. Type III hyperlipoproteinemic patients express markedly higher concentrations of apoε mRNA when compared with healthy controls. Patients with this disease are usually (>90%) homozygous for a receptor binding-defective isoform of apolipoprotein apo E (apo E2). The higher expression of apoε mRNA in the patients could, therefore, be a physiological mechanism to compensate for functionally defective apo E. The developed procedure might be valuable in assessment of apoε gene expression in human disease.

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Detection of MBL-2 gene expression in intestinal biopsies of celiac patients by in situ reverse transcription polymerase chain reaction

European Journal of Histochemistry ◽

10.4081/825 ◽

2009 ◽

Vol 47 (2) ◽

pp. 177 ◽

Cited By ~ 13

Author(s):

M Boniotto ◽

O Radillo ◽

L Braida ◽

D Pirulli ◽

A Città

Keyword(s):

Gene Expression ◽

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Chain Reaction ◽

Polymerase Chain

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Comparison of reverse transcription–quantitative polymerase chain reaction methods and platforms for single cell gene expression analysis

Analytical Biochemistry ◽

10.1016/j.ab.2012.05.010 ◽

2012 ◽

Vol 427 (2) ◽

pp. 178-186 ◽

Cited By ~ 14

Author(s):

Bridget C. Fox ◽

Alison S. Devonshire ◽

Marc-Olivier Baradez ◽

Damian Marshall ◽

Carole A. Foy

Keyword(s):

Gene Expression ◽

Polymerase Chain Reaction ◽

Single Cell ◽

Reverse Transcription ◽

Gene Expression Analysis ◽

Quantitative Polymerase Chain Reaction ◽

Chain Reaction ◽

Cell Gene Expression ◽

Polymerase Chain ◽

Cell Gene

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Differential Gene Expression Analysis of Breast Cancer by Combining Sequence-Tagged Reverse-Transcription PCR with Pyrosequencing

Springer Protocols Handbooks - Advances and Clinical Practice in Pyrosequencing ◽

10.1007/978-1-4939-3308-2_31 ◽

2016 ◽

pp. 361-369

Author(s):

Xiaodan Zhang ◽

Haiping Wu ◽

Zhiyao Chen ◽

Bingjie Zou ◽

Qinxin Song ◽

...

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Differential Gene Expression ◽

Expression Analysis ◽

Reverse Transcription ◽

Gene Expression Analysis ◽

Differential Gene Expression Analysis ◽

Reverse Transcription Pcr ◽

Differential Gene

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Study of the Alteration of Gene Expression in Adipose Tissue of Diet-Induced Obese Mice by Microarray and Reverse Transcription-Polymerase Chain Reaction Analyses

Endocrinology ◽

10.1210/en.2003-0456 ◽

2003 ◽

Vol 144 (11) ◽

pp. 4773-4782 ◽

Cited By ~ 105

Author(s):

R. C. Moraes ◽

A. Blondet ◽

K. Birkenkamp-Demtroeder ◽

J. Tirard ◽

T. F. Orntoft ◽

...

Keyword(s):

Gene Expression ◽

Polymerase Chain Reaction ◽

Adipose Tissue ◽

Reverse Transcription ◽

Obese Mice ◽

Chain Reaction ◽

Polymerase Chain

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Example of Real-Time Quantitative Reverse Transcription-PCR (Q-RT-PCR) Analysis of Bacterial Gene Expression during Mammalian Infection:Borrelia burgdorferiin Mouse Tissues

Current Protocols in Microbiology ◽

10.1002/9780471729259.mc01d03s00 ◽

2005 ◽

pp. 1D.3.1-1D.3.28 ◽

Cited By ~ 5

Author(s):

Jennifer C. Miller

Keyword(s):

Gene Expression ◽

Real Time ◽

Reverse Transcription ◽

Pcr Analysis ◽

Rt Pcr ◽

Bacterial Gene ◽

Quantitative Reverse Transcription Pcr ◽

Reverse Transcription Pcr ◽

Bacterial Gene Expression ◽

Mouse Tissues

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The contribution of increased collagen synthesis to human glomerulosclerosis: a quantitative analysis of alpha 2IV collagen mRNA expression by competitive polymerase chain reaction.

Journal of Experimental Medicine ◽

10.1084/jem.176.6.1571 ◽

1992 ◽

Vol 176 (6) ◽

pp. 1571-1576 ◽

Cited By ~ 49

Author(s):

E P Peten ◽

L J Striker ◽

M A Carome ◽

S J Elliott ◽

C W Yang ◽

...

Keyword(s):

Gene Expression ◽

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Collagen Synthesis ◽

Type Iv Collagen ◽

Chain Reaction ◽

Collagen Mrna ◽

Type Iv ◽

Polymerase Chain

We previously reported that one of the main components of the sclerotic material in human glomerular diseases was type IV collagen. In this study we examined the contribution of increased synthesis to this process at the gene expression level. Sufficient material has not been available to study type IV collagen synthesis by normal or sclerotic glomeruli in humans. We took advantage of the availability of nephrectomy specimens from patients with renal carcinoma, and of the observation that approximately 50% of these patients develop varying degrees of glomerulosclerosis. We microdissected glomeruli from 10 patients and analyzed them using in situ reverse transcription coupled with polymerase chain reaction (PCR) analyses (in situ RT-PCR). alpha 2IV collagen mRNA, after reverse transcription into cDNA, was detected in all patients and appeared to be increased in those with glomerulosclerosis (n = 5). A competitive PCR assay was developed to quantitate this change. There was an average 3.7-fold increase in glomerular type IV collagen cDNA in patients with significant sclerosis. This change was not due to an increased number of glomerular cells. Thus, glomerulosclerosis in humans is associated with an elevation of glomerular type IV collagen gene expression, suggesting that increased synthesis of type IV collagen may represent one component of this process.

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