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      Identificação de microssatélites para o mamoeiro por meio da exploração do banco de dados de DNA Translated title: Identification of microsatellites for papaya by DNA data bank exploration

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          Abstract

          Os marcadores microssatélites são ferramentas úteis em diversas análises genéticas em plantas. No caso do mamoeiro (Carica papaya L.), poucos locos de microssatélites foram descritos até o momento. Assim, o objetivo deste trabalho foi explorar a base de dados do GenBank / NCBI (National Center of Biotechnoloy Information) à procura de microssatélites de mamoeiro, visando a seu futuro uso em estudos genéticos e moleculares aplicados ao melhoramento genético. As seqüências foram obtidas no GenBank / NCBI, no formato FASTA, e analisadas para a presença de microssatélites com um mínimo de 20; 7 e 5 repetições dos motivos de mono-, di- e trinucleotídeos, respectivamente, e acima de 4 repetições para tetra- e pentanucleotídeos. Seqüências com mais de 90% de similaridade foram consideradas redundantes e, portanto, eliminadas das análises. Foram analisadas 44.591 seqüências, das quais 3.180 foram não-redundantes e apresentaram 3.947 microssatélites. Desse total, 3.587 foram classificados como microssatélites perfeitos, 8 imperfeitos, 65 interrompidos, 239 compostos-perfeitos, 8 compostos-imperfeitos e 40 compostos-interrompidos. As repetições de di- e trinucleotídeos representaram 65,7 e 14,4% do total de seqüências analisadas, respectivamente. Somente os motivos do tipo AT/TA representaram 44,1% dos microssatélites encontrados. Os motivos mais comuns de tri-, tetra- e pentanucleotídeos foram AAT, AATT e TTTAA, respectivamente. Observou-se que, nas seqüências disponíveis, o genoma do mamoeiro apresenta, em média, um microssatélite a cada 5,65 kb.

          Translated abstract

          Microsatellites markers are a useful tool in much plant genetic analysis. Currently, few microsatellites loci have been reported in Carica papaya L.. Therefore, the objective of this study was accomplishing a search for microsatellites in the public available - GenBank / NCBI (National Center of Biotechnology Information), aiming the future use in molecular and genetic studies applied to genetic breeding. The sequences were collected from GenBank / NCBI in FASTA-formatted files and analyzed for the presence of microsatellites with a minimum of 20, 7 and 5 repetitions of mono-, di- and trinucleotides, respectively, and above of 4 repetitions for tetra- and pentanucleotides. The sequences with over 90% of similarity, were considered redundant and, therefore, eliminated of the analyses. A total of 44,591 sequences had been analyzed, of which 3,180 were not redundant, showing 3,947 microsatellites. Out of those, 3,587 were classified as perfect, 8 imperfects, 65 interrupted, 239 perfect-compounds, 8 imperfect-compounds and 40 interrupted-compounds. The dinucleotides and trinucleotides repetitions represented 65.7 and 14.4% of the whole analyzed sequences, respectively. Only the AT/TA motif represented 44.1% of the microsatellites sequences. The most common motifs of tri-, tetra- and pentanucleotides were AAT, AATT and TTTAA, respectively. In the available sequences it can be observed that the papaya genome has an average one microsatellite for every 5,65 kb.

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          Evolutionary dynamics of microsatellite DNA.

          Within the past decade microsatellites have developed into one of the most popular genetic markers. Despite the widespread use of microsatellite analysis, an integral picture of the mutational dynamics of microsatellite DNA is just beginning to emerge. Here, I review both generally agreed and controversial results about the mutational dynamics of microsatellite DNA. Microsatellites are short DNA sequence stretches in which a motif of one to six bases is tandemly repeated. It has been known for some time that these sequences can differ in repeat number among individuals. With the advent of polymerase chain reaction (PCR) technology this property of microsatellite DNA was converted into a highly versatile genetic marker (Litt and Luty 1989; Tautz 1989; Weber and May 1989). Polymerase chain reaction products of different length can be amplified with primers flanking the variable microsatellite region. Due to the availability of high-throughput capillary sequencers or mass spectrography the sizing of alleles is no longer a bottleneck in microsatellite analysis. The almost random distribution of microsatellites and their high level of polymorphism greatly facilitated the construction of genetic maps (Dietrich et al. 1994; Dib et al. 1996) and enabled subsequent positional cloning of several genes. Almost at the same time, microsatellites were established as the marker of choice for the identification of individuals and paternity testing. The high sensitivity of PCR-based microsatellite analysis was not only of great benefit for forensics, but opened completely new research areas, such as the analysis of samples with limited DNA amounts (e.g., many social insects) or degraded DNA (e.g., feces, museum material) (Schlötterer and Pemberton 1998). More recently, microsatellite analysis has also been employed in population genetics (Goldstein and Schlötterer 1999). Compared with allozymes, microsatellites offer the advantage that, in principle, several thousand potentially polymorphic markers are available. Nevertheless, the application of microsatellites to population genetic questions requires a more detailed understanding of the mutation processes of microsatellite DNA as the evolutionary time frames covered in population genetics are often too long to allow novel microsatellite mutations to be ignored. Additional interest in the evolution of microsatellite DNA comes from the discovery that trinucleotide repeats, a special class of microsatellites, are involved in human neurodegenerative diseases (e.g., fragile X and Huntington's disease). A detailed understanding of the processes underlying microsatellite instability is therefore an important contribution toward a better understanding of these human neurodegenerative diseases.
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            Informativeness of human (dC-dA)n.(dG-dT)n polymorphisms.

            Abundant human interspersed repetitive DNA sequences of the form (dC-dA)n.(dG-dT)n have been shown to exhibit length polymorphisms. Examination of over 100 human (dC-dA)n.(dG-dT)n sequences revealed that the sequences differed from each other both in numbers of repeats and in repeat sequence type. Using a set of precise classification rules, the sequences were divided into three categories: perfect repeat sequences without interruptions in the runs of CA or GT dinucleotides (64% of total), imperfect repeat sequences with one or more interruptions in the run of repeats (25%), and compound repeat sequences with adjacent tandem simple repeats of a different sequence (11%). Informativeness of (dC-dA)n.(dG-dT)n markers in the perfect sequence category was found to increase with increasing average numbers of repeats. PIC values ranged from 0 at about 10 or fewer repeats to above 0.8 for sequences with about 24 or more repeats. (dC-dA)n.(dG-dT)n polymorphisms in the imperfect sequence category showed lower informativeness than expected on the basis of the total numbers of repeats. The longest run of uninterrupted CA or GT repeats was found to be the best predictor of informativeness of (dC-dA)n.(dG-dT)n polymorphisms regardless of the repeat sequence category.
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              Isolation of simple-sequence loci for use in polymerase chain reaction-based DNA fingerprinting.

              Simple sequences are short regions of tandem repetitions of mono-, di-, tri-, or tetranucleotide motifs and occur as repetitive elements in all eukaryotic genomes. These regions tend to be hypervariable in length and can therefore be exploited for DNA fingerprinting purposes, using the polymerase chain reaction with primers flanking such regions. We describe how suitable simple-sequence loci can be isolated from any given eukaryotic DNA. We show the DNA sequences for a number of variants of such loci and discuss the current results on their usefulness for DNA fingerprinting.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                rbf
                Revista Brasileira de Fruticultura
                Rev. Bras. Frutic.
                Sociedade Brasileira de Fruticultura (Jaboticabal )
                1806-9967
                September 2008
                : 30
                : 3
                : 841-845
                Affiliations
                [1 ] Embrapa Mandioca e Fruticultura Tropical
                [2 ] Embrapa Mandioca e Fruticultura Tropical
                [3 ] Embrapa Mandioca e Fruticultura Tropical
                [4 ] Universidade Federal do Recôncavo da Bahia Brazil
                Article
                S0100-29452008000300049
                10.1590/S0100-29452008000300049
                b4cf357f-a6ff-41ba-b280-ab0a01f7e8ca

                http://creativecommons.org/licenses/by/4.0/

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                Product

                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0100-2945&lng=en
                Categories
                HORTICULTURE

                Horticulture
                molecular marker,Carica papaya L.,SSR,GenBank,marcador molecular
                Horticulture
                molecular marker, Carica papaya L., SSR, GenBank, marcador molecular

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