An efficient transgenic system by TA cloning vectors and RNAi for C. elegans

Keiko Gengyo-Ando, Sawako Yoshina, Hideshi Inoue, Shohei Mitani

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

In the nematode, transgenic analyses have been performed by microinjection of DNA from various sources into the syncytium gonad. To expedite these transgenic analyses, we solved two potential problems in this work. First, we constructed an efficient TA-cloning vector system which is useful for any promoter. By amplifying the genomic DNA fragments which contain regulatory sequences with or without the coding region, we could easily construct plasmids expressing fluorescent protein fusion without considering restriction sites. We could dissect motor neurons with three colors in a single animal. Second, we used feeding RNAi to isolate transgenic strains which express lag-2::venus fusion gene. We found that the fusion protein is toxic when ectopically expressed in embryos but is functional to rescue a loss of function mutant in the lag-2 gene. Thus, the transgenic system described here should be useful to examine the protein function in the nematode.

Original languageEnglish
Pages (from-to)1345-1350
Number of pages6
JournalBiochemical and biophysical research communications
Volume349
Issue number4
DOIs
Publication statusPublished - 2006 Nov 3
Externally publishedYes

Keywords

  • Fluorescent protein
  • RNAi
  • TA cloning vector
  • Transgenic

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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