Journal of Fisheries Science and Technology

Journal of Fisheries Science and Technology

Identification,cloning and sequencing of piscidin gene from Gilthead seabream (Sparus aurata) in E. coli

Document Type : Original Research

Authors
Najme Berenjkar 1
Mohammad Reza Kalbasi 1
Saman Hoseinkhani 2
1 Department of Fisheries, Faculty of Natural Resources ans Marine Sciences, Tarbiat Modares University, Noor, Iran
2 Department of Biochemistry, Biology Sciences Faculty, Tarbiat Modares University, Tehran, Iran
Abstract
Piscidin has a wide range in killing microorganisms including bacteria, fungi, viruses, parasites and has strong anti-tumor activity and plays a role in increasing innate immunity and also does not provide resistance against bacteria; Therefore, it is of great importance in aquaculture. In this study, piscidin gene of Sparus aurata in vector pTZ57R / T was cloned. In this research, ligation product was transferred to component cell of E. coli DH5α strain. Plasmid extraction was performed from single colonies observed in ampicillin plate. Confirmation of the accuracy of single colonies grown in this research was performed by direct PCR and sequencing. The amplified cDNA fragment of the gilthead seabream piscidin gene consists of 310 nucleotides and 57 amino acids. The results of this research show that piscidin gene has been successfully cloned in pTZ57R / T vector. Comparison of nucleotide sequence of piscidin gene in this study showed high similarity with piscidin 5 of Morone chrysops. The comparison of the amino acid sequence of signal peptide piscidin is quite similar to Dicentracin-like of that species registered in the genebank, and mature peptide piscidin sequence is similar in only three amino acids to Pleorocidin-like of Poesila farmosa and Dicentracin-like of Sphaeramia orbicularis. This study could be a step towards further studies of piscidin peptide.
Keywords
piscidin peptide
Gene cloning
gilthead seabream (sparus aurata)
pTZ57R/T Vector

Subjects

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