Structure and expression of genes for proteins Rad51 and Rad51D from marine sponge Suberites domuncula

Šimatović, Ana (2009) Structure and expression of genes for proteins Rad51 and Rad51D from marine sponge Suberites domuncula. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

The proteins of interest, Rad51 and Rad51D play a very important part in maintaining mammalian genome integrity. The existence of these proteins was detected in other vertebrates, as well as in invertebrates. The model organism, marine sponge Suberites domuncula belongs to the phylum Porifera, the oldest living Metazoa. Therefore, the study of its genes/proteins that have been conserved through evolution is very interesting. Polymerase chaine reaction for multiplying DNA and Sanger dideoxynucleotide sequencing method were used in order to determine the nucleotide sequence for genes rad51 and rad51D and their intron position. Using BLAST searches of GenBank database and Clustal X and GeneDoc programmes, the obtained protein sequences were compared to their orthologs in other organisms. As expected, the protein sequences and intron position show more homology with their vertebrate orthologs than with their invertebrate orthologs. The rad51D gene was cloned into the expression vector which was used to transform BL21 cells. Insufficient protein concentration is contained in the supernatant, due to the fact that most proteins sediment and aggregate to form inclusion bodies.

Item Type: Thesis (Diploma thesis)
Keywords: sponge, Suberites domuncula, Rad51, Rad51D, introns, evolution
Supervisor: Ćetković, Helena
Date: 2009
Number of Pages: 90
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 09 Sep 2014 10:31
Last Modified: 09 Sep 2014 10:31
URI: http://digre.pmf.unizg.hr/id/eprint/2672

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