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1.
J Neurogenet ; 15(3-4): 169-78, 2001.
Artículo en Inglés | MEDLINE | ID: mdl-12092901

RESUMEN

In Drosophila melanogaster, gain-of-function mutagenesis utilizing the GAL4-UAS system has been established, allowing identification of genes that may not be easily detectable by loss-of-function screening approaches. The conditional features of misexpression systems are especially useful for studying late-stage biological processes, such as those involving adult behavior or lifespan. The gene search system, incorporating a bidirectional misexpression vector, was used to screen for genes critical for longevity determination. We have identified several genes whose misexpression in adulthood extends the fly's lifespan. Phenotypic characterization of fly lines carrying a mis-expression vector, in conjunction with obtaining information about the genomic insertion sites, creates valuable resources for the systematic functional genomics in Drosophila.


Asunto(s)
Drosophila melanogaster/genética , Genes de Insecto , Genómica , Longevidad/genética , Mutagénesis Insercional/métodos , Animales , Regulación de la Expresión Génica/genética
2.
Gene ; 253(2): 179-87, 2000 Aug 08.
Artículo en Inglés | MEDLINE | ID: mdl-10940555

RESUMEN

Microsomal glutathione S-transferase-I (MGST-I) has been thought to be important for protecting the cell from oxidative damages and/or xenobiotics. We have previously identified the Microsomal glutathione S-transferase-like (Mgstl) gene, a Drosophila homologue of human MGST-I. To investigate the function of the enzyme using Drosophila as a model system, we examined the expression pattern of Mgstl during development, and generated loss-of-function mutants to assess its in-vivo function. Mgstl was expressed in all developmental stages. It is expressed ubiquitously with the highest expression in the larval fat body, an insect organ thought to be functionally corresponding to mammalian liver, while relatively low in the central nervous system. This tissue distribution is consistent with that of MGST-I in humans or Rats. Mgstl null mutants generated from a P element insertion line showed no obvious defects in morphology, indicating that it is not essential for the development. However, their life span was significantly reduced compared to control flies, suggesting that the MGSTL protein is involved in processes somehow contributing to aging. We found an Mgstl pseudogene, which is apparently derived through the reverse transcription of Mgstl mRNA and subsequent integration into the genome.


Asunto(s)
Drosophila melanogaster/genética , Glutatión Transferasa/genética , Longevidad/genética , Animales , Secuencia de Bases , Drosophila melanogaster/embriología , Drosophila melanogaster/enzimología , Regulación del Desarrollo de la Expresión Génica , Hibridación in Situ , Datos de Secuencia Molecular , Mutagénesis , Mutación , Seudogenes/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Análisis de Supervivencia
3.
Genetics ; 151(2): 725-37, 1999 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-9927464

RESUMEN

We have constructed a P-element-based gene search vector for efficient detection of genes in Drosophila melanogaster. The vector contains two copies of the upstream activating sequence (UAS) enhancer adjacent to a core promoter, one copy near the terminal inverted repeats at each end of the vector, and oriented to direct transcription outward. Genes were detected on the basis of phenotypic changes caused by GAL4-dependent forced expression of vector-flanking DNA, and the transcripts were identified with reverse transcriptase PCR (RT-PCR) using the vector-specific primer and followed by direct sequencing. The system had a greater sensitivity than those already in use for gain-of-function screening: 64% of the vector insertion lines (394/613) showed phenotypes with forced expression of vector-flanking DNA, such as lethality or defects in adult structure. Molecular analysis of 170 randomly selected insertions with forced expression phenotypes revealed that 21% matched the sequences of cloned genes, and 18% matched reported expressed sequence tags (ESTs). Of the insertions in cloned genes, 83% were upstream of the protein-coding region. We discovered two new genes that showed sequence similarity to human genes, Ras-related protein 2 and microsomal glutathione S-transferase. The system can be useful as a tool for the functional mapping of the Drosophila genome.


Asunto(s)
Drosophila melanogaster/genética , Genes de Insecto , Análisis de Secuencia de ADN/métodos , Secuencia de Aminoácidos , Animales , Vectores Genéticos , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , Alineación de Secuencia
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