Mapping the Plasmodium falciparum genome with yeast artificial chromosomes

• Main Author: Meaney, Paul James
• Published: University of Edinburgh 1995
• Subjects: 572.8
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.657662

Yeast Artificial Chromosome cloning vectors, with their capacity to maintain up to 1 Mb of cloned DNA in a stable form, have proved extremely useful in mapping the genomes of higher eukaryotes. These vectors possess features which can circumvent some of the problems associated with classical molecular manipulation in <I>Plasmodium falciparum. </I>The research presented in this thesis is aimed at contributing to genome mapping in <I>P. falciparum. </I>The primary objective is the construction of a complete, detailed YAC- based physical map of chromosome 6, with a resolution of 10 Kb. To accomplish this, an 1100 YAC library of the <I>P. falciparum </I>isolate HB3 was constructed. The library contains clones with an average insert size of 100 Kb. Insert DNA is stable when cultured over 100 generations and the library is predicted to have a 4/5 fold genome redundancy, corresponding to 90% of the genome. Chromosome 6 specific YAC's have been isolated and three contigs initiated. Overlapping YAC's have been identified by using Sequence Tagged Site markers obtained from the 5 and 3 ends of each YAC by Inverse PCR. A total of 700 Kb of <I>P. falciparum </I>DNA has been cloned and this has been extensively mapped with seven restriction enzyme. Maps for all available YAC's will be presented. In addition, an attempt has been made to evaluate the degree of stage specific gene expression of cloned DNA within each YAC. The implications of these findings for genome mapping in <I>P. falciparum</I> will be discussed in the thesis.