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      Genomic organization and expression profile of the mucin-associated surface protein ( masp) family of the human pathogen Trypanosoma cruzi

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          Abstract

          A novel large multigene family was recently identified in the human pathogen Trypanosoma cruzi, causative agent of Chagas disease, and corresponds to ∼6% of the parasite diploid genome. The predicted gene products, mucin-associated surface proteins (MASPs), are characterized by highly conserved N- and C-terminal domains and a strikingly variable and repetitive central region. We report here an analysis of the genomic organization and expression profile of masp genes. Masps are not randomly distributed throughout the genome but instead are clustered with genes encoding mucin and other surface protein families. Masp transcripts vary in size, are preferentially expressed during the trypomastigote stage and contain highly conserved 5′ and 3′ untranslated regions. A sequence analysis of a trypomastigote cDNA library reveals the expression of multiple masp variants with a bias towards a particular masp subgroup. Immunofluorescence assays using antibodies generated against a MASP peptide reveals that the expression of particular MASPs at the cell membrane is limited to subsets of the parasite population. Western blots of phosphatidylinositol-specific phospholipase C (PI-PLC)-treated parasites suggest that MASP may be GPI-anchored and shed into the medium culture, thus contributing to the large repertoire of parasite polypeptides that are exposed to the host immune system.

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          Molecular Cloning : A Laboratory Manual

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            Consed: a graphical tool for sequence finishing.

            Sequencing of large clones or small genomes is generally done by the shotgun approach (Anderson et al. 1982). This has two phases: (1) a shotgun phase in which a number of reads are generated from random subclones and assembled into contigs, followed by (2) a directed, or finishing phase in which the assembly is inspected for correctness and for various kinds of data anomalies (such as contaminant reads, unremoved vector sequence, and chimeric or deleted reads), additional data are collected to close gaps and resolve low quality regions, and editing is performed to correct assembly or base-calling errors. Finishing is currently a bottleneck in large-scale sequencing efforts, and throughput gains will depend both on reducing the need for human intervention and making it as efficient as possible. We have developed a finishing tool, consed, which attempts to implement these principles. A distinguishing feature relative to other programs is the use of error probabilities from our programs phred and phrap as an objective criterion to guide the entire finishing process. More information is available at http:// www.genome.washington.edu/consed/consed. html.
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              The genome of the African trypanosome Trypanosoma brucei.

              African trypanosomes cause human sleeping sickness and livestock trypanosomiasis in sub-Saharan Africa. We present the sequence and analysis of the 11 megabase-sized chromosomes of Trypanosoma brucei. The 26-megabase genome contains 9068 predicted genes, including approximately 900 pseudogenes and approximately 1700 T. brucei-specific genes. Large subtelomeric arrays contain an archive of 806 variant surface glycoprotein (VSG) genes used by the parasite to evade the mammalian immune system. Most VSG genes are pseudogenes, which may be used to generate expressed mosaic genes by ectopic recombination. Comparisons of the cytoskeleton and endocytic trafficking systems with those of humans and other eukaryotic organisms reveal major differences. A comparison of metabolic pathways encoded by the genomes of T. brucei, T. cruzi, and Leishmania major reveals the least overall metabolic capability in T. brucei and the greatest in L. major. Horizontal transfer of genes of bacterial origin has contributed to some of the metabolic differences in these parasites, and a number of novel potential drug targets have been identified.
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                Author and article information

                Journal
                Nucleic Acids Res
                Nucleic Acids Res
                nar
                nar
                Nucleic Acids Research
                Oxford University Press
                0305-1048
                1362-4962
                June 2009
                31 March 2009
                31 March 2009
                : 37
                : 10
                : 3407-3417
                Affiliations
                1Department of Parasitology, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil, 2Department of Cell Biology and Molecular Genetics, 3Center for Bioinformatics and Computational Biology, 4Maryland Pathogen Research Institute, University of Maryland, College Park, MD 20742, USA, 5Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil and 6J. Craig Venter Institute, Rockville, MD 20850, USA
                Author notes
                *To whom correspondence should be addressed. Tel: +1 301 4052999; Fax: +1 301 838 0208; Email: elsayed@ 123456umd.edu
                Correspondence may also be addressed to Daniella C. Bartholomeu. Tel: +5531 3409 2825; Fax: +5531 3409 2970; Email: daniella@ 123456icb.ufmg.br
                Article
                gkp172
                10.1093/nar/gkp172
                2691823
                19336417
                7d01bf35-ed5c-4b84-a983-04488554e4b4
                © 2009 The Author(s)

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 9 January 2009
                : 27 February 2009
                : 2 March 2009
                Categories
                Genomics

                Genetics
                Genetics

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