Molecular Cloning and Phylogenetic Analysis of a Chitin Deacetylase Isolated from the Epidermis of the Red Snow Crab Chionoecetes japonicas - Advances in Bioscience and Biotechnology

ABB > Vol.9 No.1, January 2018

Molecular Cloning and Phylogenetic Analysis of a Chitin Deacetylase Isolated from the Epidermis of the Red Snow Crab Chionoecetes japonicas ()

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DOI: 10.4236/abb.2018.91005 941 Downloads 1,895 Views Citations

Author(s)

Kakeru Fujimori, Hideto Fukushima, Masahiro Matsumiya

Affiliation(s)

Department of Marine Science and Resources, College of Bioresource Sciences, Nihon University, Fujisawa, Japan.

ABSTRACT

Chitin deacetylase (CDA; EC 3. 5. 1. 41) catalyzes the deacetylation of chitin. In this study, we successfully cloned and sequenced a chitin deacetylase gene from the red snow crab Chionoecetes japonicas. By using reverse transcription-polymerase chain reaction (RT-PCR) and 5' and 3' rapid amplification of cDNA ends, we obtained a 2141-bp amplicon containing a chitin deacetylase gene (CjCDA) from the epidermis of C. japonicas. The amplicon contains a 1575-bp open reading frame that is predicted to encode a 525-amino acid protein. The structure predicted from the deduced amino acid sequence included an N-terminal signal peptide, chitin-binding domain (CBD), low-density lipoprotein receptor class A domain (LDL-A), and catalytic domain. Comparative analysis of the deduced amino acid sequence of CjCDA revealed the highest homology (74%) to gastrolith protein 59 of Cherax quadricarinatus. We used RT-PCR to evaluate the expression of CjCDA in various tissues of C. japonicas, and we observed that CjCDA was expressed only in the epidermis. A phylogenetic analysis, using the amino acid sequences of CjCDA and other known chitin deacetylases, showed that CjCDA belonged to a group of crustacean chitin deacetylases. To our knowledge, this is the first study reporting the cDNA cloning of a chitin deacetylase from a crab.

KEYWORDS

Chitin Deacetylase, Molecular Cloning, Chionoecetes japonicas, Phylogenetic Analysis, Expression Analysis

Share and Cite:

Fujimori, K. , Fukushima, H. and Matsumiya, M. (2018) Molecular Cloning and Phylogenetic Analysis of a Chitin Deacetylase Isolated from the Epidermis of the Red Snow Crab Chionoecetes japonicas. Advances in Bioscience and Biotechnology, 9, 52-62. doi: 10.4236/abb.2018.91005.

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