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Title:	Molecular cloning and characterisation of ferritin in the asian seabass (Lates calcarifer)
Authors:	Lee Jun Hoe (P45167)
Supervisor:	Wan Kiew Lian, Prof Dr
Keywords:	Ferritin Molecular aspects Giant perch Clones Carrier proteins
Issue Date:	8-Mar-2012
Description:	The Asian seabass (Lates calcarifer) is one of the major marine aquaculture fish species in Malaysia. One of the major challenges to the L. calcarifer aquaculture industry is the management of diseases caused by pathogens. Ferritin is a highly-conserved protein that functions to sequester iron and appears to be upregulated in infected L. calcarifer. In mammals, two ferritin subunits known as the heavy (H) and light (L) chains have been well-characterised, but there is little information on teleost ferritins. The objectives of this study are to generate the full-length cDNA sequences of ferritin from L. calcarifer and to characterise them through phylogenetic analysis and expression studies. Two full-length putative ferritin cDNA sequences, Latca_M and Latca_H, were generated through the 5' and 3'-RACE. Latca_H and Latca_M consisted of 1130 and 1148 nucleotides, respectively and sequence analysis indicated the presence of a conserved iron-responsive element in the 5' end of the untranslated region of both sequences. A comparison between the gene and cDNA sequences also showed that both Latca_H and Latca_M consist of a 4 exon/3 intron gene structure, which is conserved in other vertebrate ferritins. Multiple sequence alignment showed that Latca_H possesses all the conserved ferroxidase residues found in H-chains, while Latca_M not only possesses the ferroxidase residues, but also the nucleation residues found in L-chains. Phylogenetic analysis showed that Latca_M and Latca_H are clustered with the teleost ferritin middle (M) and H-chains, respectively. Additionally, the teleost M-chains are found to be orthologous to the L-chains in mammals. Analysis of quantitative real-time PCR revealed that Latca_M and Latca_H were differentially expressed in the liver, spleen, gills, and kidney after infection with Aeromonas hydrophila, a main pathogen in aquaculture. It was found that the expression of Latca_H was consistently higher than Latca_M in all tissues with the exception of kidney. These results suggest that Latca_H is capable of sequestering iron at a faster rate compared to Latca_M. However, the expression of Latca_M was found to be downregulated in the liver. This was unexpected and suggest the possibility of undiscovered aspects of the iron regulation mechanism during infection. Results from this study showed the significant roles of ferritin in fish immunity. Further studies on the regulation of the ferritin chains and their interaction with other components of the immune system will contribute towards a more comprehensive understanding of the host-pathogen interaction in fish.,Tesis ini tiada perakuan deklarasi pelajar,Sarjana Sains
Pages:	130
Call Number:	QP552.F47L434 2012 tesis
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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