Pembinaan dan Pencirian Perpustakaan Genom Siakap (Lates calcarifer)

Sea bass (Lates calcarifer) is an economically importantfish in Malaysia. In this study, a genomic library of L. calcarifer was constructed. Genomic DNA was isolated from the muscle tissue of sea bass, sheared by nebulisation, blunt-end repaired and ligated into the pCR4Blunt-TOPO plasmid vector. The ligation mixture was then transformed into Escherichia coli. In order to characterize the constructed genomic library, plasmid DNA was extracted from randomly selected recombinant clones. Gel electrophoresis showed that these recombinant clones contain inserts with a size range of 0.5 kb - 3.5 kb, with an average size of 1.7 kb. DNA sequencing was then carried out using the extracted DNA plasmid as template, and a total of 121 random sequences were generatedfrom the L. calcarifer genomic library. Analysis of these sequences implies that the library is representative of L. calcarifer genome. Based on sequence similarity database searches, 17% of the random sequences were found to have significant BLAST hits, indicating that they represent L. calcarifer genes. Analysis using the RepeatMasker program reveals that 35% of the random sequences were identified as repeat elements. Initial characterization of this genomic library indicates that it is a potentially useful resource in the study of the L. calcarifer genome.