Genetic cloning and characterization of Spalax ehrenbergi mole rat H3 histone gene

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Genetic cloning and characterization of Spalax ehrenbergi mole rat H3 histone gene

 

Author: Leung, Fo-man
Title: Genetic cloning and characterization of Spalax ehrenbergi mole rat H3 histone gene
Degree: M.Sc.
Year: 2001
Subject: Spalacidae
Genetics
Cloning
Hong Kong Polytechnic University -- Dissertations
Department: Multi-disciplinary Studies
Dept. of Applied Biology and Chemical Technology
Pages: vii, 45 leaves : ill., 1 map ; 30 cm
Language: English
InnoPac Record: http://library.polyu.edu.hk/record=b1551199
URI: http://theses.lib.polyu.edu.hk/handle/200/60
Abstract: Subterranean mole rats are species of Spalax ebrenbergi in the genus of Spalax under Spalacidae family, which are commonly inhabited in Israel and Eastern Europe in a sealed subterraneous environment. Recent studies of species S. ehrenbergi in Israel have revealed that S ehrenbergi is in fact comprised of four morphologically indistinguishable chromosomal species (2n=52, 54, 58 and 60). Interestingly, the distributions of four chromosomal species are found related to the climatic regimes, of which, increasing diploid chromosome numbers radiate toward from dry-cool to humid-warm environments in Israel. S. ehrenbergi provides a good model for studying evolutionary speciation and climatic adaptive radiation. Speciation is caused by the prolonged accumulation of genetic changes during the history of evolution. In order to understand speciation and its role in evolution, it is useful to know how much genetic change takes place during the course of species development. The question of how much genetic change during speciation has become answerable only for recent development of genetic characterizations. Histone genes, being one of the most conserved genes and performing same function among al1 the eukaryotic organisms, are ideal genes for studying speciation and adaptive radiation. In this project, histone H3 gene of eight individuals of different populations from four chromosomal species (2n=52, 54, 58 and 60) of Spalax ehrenbergi were obtained by Polymerase Chain Reaction (PCR) amplification from their genomic DNA. In order to design respective pair of primers for such purpose, known histone nucleotide sequences of human, mouse, rat, chicken and duck were downloaded from the NCBI genebank. Those sequences were then aligned by ClustalX software based on the best similarity of nucleotides among the species under test, the common conserved oligonucleotides fall within the consensus region at both ends of the histone H3 gene were considered as the potential primer candidates, they were chosen as the forward and reverse primer. The primers were successfully amplified 520bps intact histone H3 genes of eight populations of Spa/ax chrenbergi. Those purified PCR derived histone H3 genes were finally cloned into the pBluescript II plasniid vector SK+. Subsequently, DNA sequencing to characterize histone H3 genes by fluorescent labeled primers was performed, but only partial sequences had been obtained.

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