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United States Department of Agriculture

Agricultural Research Service

Research Project: MOLECULAR ANALYSIS OF VIRULENCE DETERMINANTS OF SELECT BACTERIA IN FISH DISEASES Title: Cloning and Sequencing of Channel Catfish (Ictalurus Punctatus) Matrix Metalloproteinase-9

Authors
item Yeh, Hung-Yueh
item Klesius, Phillip

Submitted to: Genbank
Publication Type: Other
Publication Acceptance Date: June 30, 2006
Publication Date: August 10, 2006
Citation: Yeh, H., Klesius, P.H. 2006. Cloning and sequencing of channel catfish (Ictalurus punctatus) matrix metalloproteinase-9. Genbank.

Technical Abstract: In the course of studying pathogenesis of enteric septicemia of catfish, we noted that channel catfish matrix metalloproteinase-9 (MMP-9) gene was up-regulated after Edwardsiella ictaluri infection. In this study, we cloned and sequenced using the RACE. The complete sequence of the CC MMP-9 cDNA gene consisted of 2562 nucleotides. Analysis of the nucleotide sequence revealed one open reading frame and 5’- and 3’-end untranslated regions. The 5’-end untranslated region had 103 bases that are shorter than the corresponding region of rainbow trout (Oncorhynchus mykiss) (Johnson et al., 2004). The 3’-end untranslated region had 398 bases in length. As seen in mammalian and other fish mRNAs, the CCMMP-9 mRNA instability motifs (ATTTA) that regulate mRNA degradation (Zubiaga et al., 1995) were located within the 3’-end untranslated region. In addition, a polyadenylation signal sequence (AATAAA) was located 24 bp upstream from the polyadenylation tail. The open reading frame potentially encoded a 686-amino-acid peptide with a calculated molecular mass (without glycosylation) of approximate 77.4 kDa. In addition, analyzed by the SignalIP 3.0 program (Bendtsen et al., 2004), the CCMMP-9 included a signal peptide with a cleavage site at positions 20 and 21 (AWS-HP). Like mammalian MMP-9, CCMMP-9 analyzed by the NetOGlyc 3.1 program (Julenius et al., 2005) had potentially heavy O-glycosylation sites between positions 447 and 490. More than ten clones were sequenced and the same results were obtained.

Last Modified: 10/24/2014
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