Cloning, Phylogenetic Analysis and 3D Modeling of a Putative Lysosomal Acid Lipase from the Camel, Camelus dromedariusReport as inadecuate




Cloning, Phylogenetic Analysis and 3D Modeling of a Putative Lysosomal Acid Lipase from the Camel, Camelus dromedarius - Download this document for free, or read online. Document in PDF available to download.

1

Department of Biochemistry, College of Science, King Saud University, Bld #5, Room 2A26, P.O. Box 2455, Riyadh 11451, Saudi Arabia

2

Department of Molecular Biology, Genetic Engineering Division, National Research Center, Dokki, Cairo 12311, Egypt





Abstract Acid lipase belongs to a family of enzymes that is mainly present in lysosomes of different organs and the stomach. It is characterized by its capacity to withstand acidic conditions while maintaining high lipolytic activity. We cloned for the first time the full coding sequence of camel’s lysosomal acid lipase, cLIPA using RT-PCR technique Genbank accession numbers JF803951 and AEG75815, for the nucleotide and aminoacid sequences respectively. The cDNA sequencing revealed an open reading frame of 1,197 nucleotides that encodes a protein of 399 aminoacids which was similar to that from other related mammalian species. Bioinformatic analysis was used to determine the aminoacid sequence, 3D structure and phylogeny of cLIPA. Bioinformatics analysis suggested the molecular weight of the translated protein to be 45.57 kDa, which could be decreased to 43.16 kDa after the removal of a signal peptide comprising the first 21 aminoacids. The deduced cLIPA sequences exhibited high identity with Equus caballus 86%, Numascus leucogenys 85%, Homo sapiens 84%, Sus scrofa 84%, Bos taurus 82% and Ovis aries 81%. cLIPA shows high aminoacid sequence identity with human and dog-gastric lipases 58%, and 59% respectively which makes it relevant to build a 3D structure model for cLIPA. The comparison confirms the presence of the catalytic triad and the oxyanion hole in cLIPA. Phylogenetic analysis revealed that camel cLIPA is grouped with monkey, human, pig, cow and goat. The level of expression of cLIPA in five camel tissues was examined using Real Time-PCR. The highest level of cLIPA transcript was found in the camel testis 162%, followed by spleen 129%, liver 100%, kidney 20.5% and lung 17.4%.

Keywords: LAL-LIPA; 3D modeling; cloning; phylogenetic tree; qPCR; one-humped camel LAL-LIPA; 3D modeling; cloning; phylogenetic tree; qPCR; one-humped camel





Author: Farid Shokry Ataya 1,2

Source: http://mdpi.com/



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