Detection of Differentially Expressed Genes in an Isogenic Breast Metastasis Model using RNA Arbitrarily Primed-Polymerase Chain Reaction Coupled with Array Hybridization (RAP-Array)

The first adult-repopulating hematopoietic stem cells (HSCs) emerge in the mouse aorta-gonad-mesonephros (AGM) region at embryonic day 10.5 prior to their appearance in the yolk sac and fetal liver. Although several genes are implicated in the regulation of HSCs, there are gaps in our understanding of the processes taking place in the AGM at the time of HSC emergence. To identify genes involved in AGM HSC emergence, we performed differential display reverse transcriptase–polymerase chain reaction (DD RT-PCR). Differentially expressed genes included β-catenin and homologs of human TM9SF2 and TAB2. We characterized the expression pattern of Wnt/β-catenin signaling,mTM9SF2, and mTAB2 in the embryo and adult. Interestingly, the expression of mouse TAB2 (mTAB2) in the E11 dorsal aorta endothelium suggests a role for mTAB2 in HSC emergence and/or regulation. The identification of differentially expressed genes in the AGM region should yield further insights into the development of this tissue and into the emergence and regulation of HSCs.

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Identification of differentially expressed genes in rheumatoid arthritis by a combination of complementary DNA array and RNA arbitrarily primed-polymerase chain reaction

Arthritis & Rheumatism ◽

10.1002/1529-0131(200201)46:1<52::aid-art10048>3.0.co;2-1 ◽

2002 ◽

Vol 46 (1) ◽

pp. 52-63 ◽

Cited By ~ 39

Author(s):

E. Neumann ◽

F. Kullmann ◽

M. Judex ◽

H. P. J�sten ◽

D. Wessinghage ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Polymerase Chain Reaction ◽

Differentially Expressed Genes ◽

Differentially Expressed ◽

Dna Array ◽

Chain Reaction ◽

Complementary Dna ◽

Polymerase Chain

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RAP-PCR fingerprinting reveals time-dependent expression of development-related genes following differentiation process of Bacillus thuringiensis

Canadian Journal of Microbiology ◽

10.1139/cjm-2015-0212 ◽

2015 ◽

Vol 61 (9) ◽

pp. 683-690 ◽

Cited By ~ 1

Author(s):

Tianpei Huang ◽

Xiaomin Yu ◽

Ivan Gelbič ◽

Xiong Guan

Keyword(s):

Polymerase Chain Reaction ◽

Bacillus Thuringiensis ◽

Molecular Mechanisms ◽

Tricarboxylic Acid Cycle ◽

Expression Profiles ◽

Differentially Expressed ◽

Differentiation Process ◽

Chain Reaction ◽

Quinone Oxidoreductase ◽

Polymerase Chain

Gene expression profiles are important data to reveal the functions of genes putatively involved in crucial biological processes. RNA arbitrarily primed polymerase chain reaction (RAP-PCR) and specifically primed reverse transcription polymerase chain reaction (RT-PCR) were combined to screen differentially expressed genes following development of a commercial Bacillus thuringiensis subsp. kurstaki strain 8010 (serotype 3a3b). Six differentially expressed transcripts (RAP1 to RAP6) were obtained. RAP1 encoded a putative triple helix repeat-containing collagen or an exosporium protein H related to spore pathogenicity. RAP2 was homologous to a ClpX protease and an ATP-dependent protease La (LonB), which likely acted as virulence factors. RAP3 was homologous to a beta subunit of propionyl-CoA carboxylase required for the development of Myxococcus xanthus. RAP4 had homology to a quinone oxidoreductase involved in electron transport and ATP formation. RAP5 showed significant homology to a uridine kinase that mediates phosphorylation of uridine and azauridine. RAP6 shared high sequence identity with 3-methyl-2-oxobutanoate-hydroxymethyltransferase (also known as ketopantoate hydroxymethyltransferase or PanB) involved in the operation of the tricarboxylic acid cycle. The findings described here would help to elucidate the molecular mechanisms underlying the differentiation process of B. thuringiensis and unravel novel pathogenic genes.

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Identification of a novel DNA sequence differentially expressed between normal human CD34+CD38hi and CD34+CD38lo marrow cells

Blood ◽

10.1182/blood.v86.2.548.bloodjournal862548 ◽

1995 ◽

Vol 86 (2) ◽

pp. 548-556 ◽

Cited By ~ 23

Author(s):

L Graf ◽

B Torok-Storb

Keyword(s):

Bone Marrow ◽

Polymerase Chain Reaction ◽

Dna Sequence ◽

Organ Donor ◽

Radioactive Tracer ◽

Differentially Expressed ◽

Chain Reaction ◽

Relative Level ◽

Human Organ ◽

Polymerase Chain

We have applied the recently developed differential display method to extend the molecular characterization of the less mature CD34+CD38lo bone marrow progenitors in comparison with the CD34+CD38hi cells to better understand their functional differences. Immunomagnetic enrichment of CD34+ cells followed by flow cytometry was used to isolate CD34+CD38lo and CD34+CD38hi cells from human organ donor bone marrow. A limited set of the poly A+ RNA sequences present in these two cell populations was amplified by a combination of reverse transcription with an anchored oligo dT-based primer and polymerase chain reaction with the same oligo dT primer and arbitrary decamers. A radioactive tracer allowed these sequences to be displayed as a series of bands on a denaturing polyacrylamide gel. Eight bands were chosen that appeared in multiple displays to represent gene sequences differentially expressed between CD34+CD38hi and CD34+CD38lo cells. Comparison of the sequences with public DNA sequence databases available identified one sequence as myeloperoxidase. Two other clones matched sequence fragments of unknown function, whereas the remaining five are novel sequences not present in existing databases. The relative level of expression of all of the sequences was tested by an independent reverse transcriptase-polymerase chain reaction with sequence-specific oligonucleotide primers. The lower level of myeloperoxidase mRNA in CD34+CD38lo cells was confirmed, as was the higher expression of the novel sequence 345. Sequence 345 expression is highest in CD34+CD38- cells and decreases with increased CD38 expression. It is expressed in negligible amounts in hematopoietic cell lines and other sources of human tissue, suggesting it may have a functional role in normal hematopoiesis.

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