An Efficient Transgenic System by Ta Cloning Vectors and Rnai for C. Elegans.
From: Department of Physiology, Tokyo Women’s Medical University School of Medicine, Tokyo 162-8666, Japan.
Biochemical and biophysical research communications
- Publish Date: Nov 2006
- ISSN: 0006-291X
- Volume: 349
- Issue: 4
- Pages: 1345-50
- Medium: Print
- Language: English
- Citation (JAMA): Gengyo-Ando Keiko, Yoshina Sawako, Inoue Hideshi, et al. An Efficient Transgenic System by Ta Cloning Vectors and Rnai for C. Elegans.. Biochem. Biophys. Res. Commun. Nov 2006;349:1345-50
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.
Mesh Headings (Keywords): Animals, Animals, Genetically Modified, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Cloning, Molecular, Genetic Vectors, Polymerase Chain Reaction, RNA Interference, Recombinant Fusion Proteins, Transfection
Check for Full Text / PubMed Unique Identifier (PMID): 16979594
This abstract is part of PubMed, a service of the U.S. National Library of Medicine. PubMed includes more than 17 million citations from MEDLINE and other life science journals for biomedical articles. See Copyright and Disclaimers.
Linked medical terms appearing on this page are added by Healia to help readers find more information and are not part of the original PubMed document.
The data herein was last updated on July 8th, 2008 and may not reflect the most current and accurate data available from NLM.