The gene search system: A method for efficient detection and rapid molecular identification of genes in Drosophila melanogaster

Gakuta Toba, Takashi Ohsako, Naomasa Miyata, Tsuyoshi Ohtsuka, Ki Hyeon Seong, Toshiro Aigaki

    Research output: Contribution to journalArticlepeer-review

    203 Citations (Scopus)

    Abstract

    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 activation 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.

    Original languageEnglish
    Pages (from-to)725-737
    Number of pages13
    JournalGenetics
    Volume151
    Issue number2
    Publication statusPublished - 1999 Feb 1

    ASJC Scopus subject areas

    • Genetics

    Fingerprint Dive into the research topics of 'The gene search system: A method for efficient detection and rapid molecular identification of genes in Drosophila melanogaster'. Together they form a unique fingerprint.

    Cite this