cDNA Cloning and Sequencing, Gene Expression, and Immunolocalization of Thrombomodulin in the Sprague-Dawley Rat (original) (raw)

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Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

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Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

Search for other works by this author on:

,

Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

Search for other works by this author on:

,

Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

Search for other works by this author on:

,

Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

Search for other works by this author on:

,

Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

Search for other works by this author on:

Departments of Surgery and Pathology, University of Arkansas for Medical Sciences and John L. McClellan VAMC

4301 W. Markham, Little Rock, AR 72205, USA

* To whom correspondence should be addressed. Arkansas Cancer Research Center 4301 West Markham, Slot 725 Little Rock, AR 72205,USA. Tel. +1-501-686-7912, Fax. +1-501-686-7861, E-mail: mhjensen@life.uams.edu

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Received:

18 September 1998

Revision received:

14 October 1998

Published:

01 February 1999

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Junru Wang, Aiwei Yao, Jing-Yi Wang, Ching-Ching Sung, Louis M. Fink, James W. Hardin, Martin Hauer-Jensen, cDNA Cloning and Sequencing, Gene Expression, and Immunolocalization of Thrombomodulin in the Sprague-Dawley Rat, DNA Research, Volume 6, Issue 1, 1999, Pages 57–62, https://doi.org/10.1093/dnares/6.1.57
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Abstract

Thrombomodulin (TM), in addition to its significance in the protein C anticoagulant pathway and cardiovascular diseases, has recently been shown to play important roles in normal embryonic development, several inflammatory conditions, as well as in tumor biology and in the pathogenesis of chronic radiation toxicity. We cloned and sequenced the cDNA encoding the complete TM protein from the Sprague-Dawley rat. The cDNA sequence consisted of a 78-bp 5′ non-coding region and a 1731-bp open reading frame encoding 577 amino acids. Comparison of the deduced amino acid sequences showed Sprague-Dawley rat TM to be 87% homologous with mouse and 70.3% with human TM. In addition to the previously described highly conserved region in the lectin-like domain, another region was found which possessed significant homology among the species and may be involved in regulating cell surface expression of TM. Primers and fluorogenic probe for 5′ exonuclease-based real time RT-PCR detection (TaqMan™ PCR) were constructed based on the cDNA sequence information and used to determine steady-state TM mRNA levels in lung, intestine, kidney, brain, and liver. The highest TM mRNA levels were found in lung and the lowest in liver. Immunohistochemistry confirmed that TM was mainly localized on the endothelium of blood vessels and lymphatics. The alveolar capillaries of lung showed the strongest immunoreactivity, whereas the endothelium of hepatic sinusoids and cerebral cortex were virtually negative.

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