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Int. J. Mol. Sci. 2015, 16(5), 9922-9935; doi:10.3390/ijms16059922

Quantitative Real-Time PCR Analysis of YKL-40 and Its Comparison with Mammalian Chitinase mRNAs in Normal Human Tissues Using a Single Standard DNA

1
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan
2
Research Fellow of Japan Society for the Promotion of Science (DC2), Koujimachi, Chiyoda-ku, Tokyo 102-0083, Japan
3
Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Ritva Tikkanen
Received: 11 March 2015 / Revised: 22 April 2015 / Accepted: 22 April 2015 / Published: 30 April 2015
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

YKL-40 (YKL for the first three N-terminal residues of a 40 kDa protein) belongs to a group of human chitinase-like proteins (CLPs), which are similar to chitinases but lack chitinolytic activity. YKL-40 mRNA and its protein levels have been reported elevated in multiple disorders including asthma, cystic fibrosis, rheumatoid arthritis and malignant tumors. Here, we quantified the YKL-40 mRNA levels and compared them with chitinases and housekeeping genes in normal human tissues. To establish the quantitative real-time PCR (qPCR) system for evaluation of relative YKL-40 mRNA levels, we constructed a human standard DNA molecule by ligating cDNAs of YKL-40, two mammalian chitinases and two housekeeping genes in a one-to-one ratio. We generated cDNAs from various normal human tissues and analyzed the YKL-40 mRNA expression levels using a qPCR system with the standard DNA. We found that YKL-40 mRNA is present widely in human tissues while its expression patterns exhibit clear tissue specificity. Highest YKL-40 mRNA levels were detected in the liver, followed by kidney, trachea and lung. The levels of YKL-40 mRNA in the kidney and liver were more than 100-times higher than those of chitotriosidase mRNA. Our study provides for the first time a comprehensive analysis of the relative expression levels of YKL-40 mRNA versus mammalian chitinases in normal human tissues. View Full-Text
Keywords: asthma; chitinase; chitinase-like protein; cystic fibrosis; gene expression analysis; malignant tumors; normal human tissues; quantitative real-time PCR system; rheumatoid arthritis; YKL-40 asthma; chitinase; chitinase-like protein; cystic fibrosis; gene expression analysis; malignant tumors; normal human tissues; quantitative real-time PCR system; rheumatoid arthritis; YKL-40
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Ohno, M.; Bauer, P.O.; Kida, Y.; Sakaguchi, M.; Sugahara, Y.; Oyama, F. Quantitative Real-Time PCR Analysis of YKL-40 and Its Comparison with Mammalian Chitinase mRNAs in Normal Human Tissues Using a Single Standard DNA. Int. J. Mol. Sci. 2015, 16, 9922-9935.

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