Navigation Links
Big Blue Rodent Cell Cultures for Mutagenesis Studies

Rat and mouse embryonic cell lines are useful for in vitro mutagenesis studies

Denise L. Wyborski
Stratagene Cloning Systems, Inc.

The Big Blue rat cell line and the Big Blue transgenic mouse embryonic cell line are useful for in vitro mutagenesis studies. These cell lines are part of the Big Blue transgenic rodent mutagenesis assay system,* a system for detecting mutations from several known mutagens in a variety of tissues from transgenic mice and rats.1-4

figure 1

The Big Blue rat cell line is derived from a Rat2 embryonic fibroblast cell line transfected with the Big Blue lambda shuttle vector and the pSV2Neo plasmid, which provides an antibiotic-selection marker.5-7 The Big Blue lambda shuttle vector (figure 1) is 45 kb in size and contains the lacI and alphalacZ genes. Each cell contains 50 to 70 copies of the lambda shuttle vector integrated into the genome at two sites. The Big Blue rat cell line is resistant to the antibiotic G418 and is a thymidine kinase mutant; the chromosome number is polyploid. The spontaneous mutant frequency of this cell line is approximately 3.7 x 105.

The Big Blue transgenic mouse embryonic cell line is derived from embryonic cells of transgenic mice containing the Big Blue lambda shuttle vector. The cell line was immortalized by chemical transformation and differentiation and has survived more than 50 generations. Each cell contains at least 40 copies of the Big Blue lambda shuttle vector integrated into the genome of the cell at the identical location as in the genome of the transgenic mice. This cell line exhibits a spontaneous mutant frequency of approximately 9.28 x 105.

The Big Blue cell lines use the lac I gene of the Big Blue lambda shuttle vector as the target for mutagenesis. After the cells are treated with a test compound, in vitro packaging rapidly and efficiently recovers the lambda shuttle vector from the genomic DNA of the cell. These cell lines exhibit rescue efficiencies between 5x104 and 1x105 plaque-forming units per g of genomic DNA. Bacterial detection systems screen for mutations in the lacI target gene. The target gene can then be recovered from mutant plaques for sequence analysis.

In addition to short-term in vitro mutagenesis studies, these cell lines are useful for various toxicological applications, studies of the effects of oncogenes and tumor suppressor genes on mutation frequencies and studies of DNA repair.

REFERENCES
  1. Kohler, S.W., et al. (1991) Proc. Natl. Acad. Sci. USA 88: 78597962.

  2. Dycaico, M.J., et al. (1994) Mutat. Res. 307: 461478.

  3. Rogers, B.J., et al. (1995) Mutat. Res. 327: 5766.

  4. Dycaico, M.J., et al. (1996) Carcinogenesis 17: 23472356.

  5. Wyborski, D.L., Malkhosyan, S., Moores, J.C., Dycaico, M.J., and Short, J.M. (1994) Strategies 7: 5556.

  6. Wyborski, D.L., et al. (1995) Mutat. Res. 334: 161165.

  7. Manjanatha, M.G., et al. (1996) Environ. Mol. Mutagen. 27: 44


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment
2. High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
3. Randomize Gene Sequences with New PCR Mutagenesis Kit
4. In-Vitro Metabolism Studies Using Data-Dependent LC/MSn
5. Precursor miRNAs for Successful miRNA Functional Studies
6. DGGE Analysis for Non-Invasive Studies of Primate Diet
7. Using the DCode System to Identify DNA Sequence Variation for Studies of Population Structure in Marine Organisms
8. SNPs Powerful Tools for Association Studies
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... TORONTO , June 23, 2016 /PRNewswire/ - ... Ontario biotechnology company, Propellon ... the development and commercialization of a portfolio of ... cancers. Epigenetic targets such as WDR5 represent an ... contribute significantly in precision medicine for cancer patients. ...
(Date:6/23/2016)... ... June 23, 2016 , ... Charm Sciences, Inc. is pleased ... received AOAC Research Institute approval 061601. , “This is another AOAC-RI approval of ... Salter, Vice President of Regulatory and Industrial Affairs. “The Peel Plate methods perform ...
(Date:6/23/2016)... , June 23, 2016   EpiBiome , ... secured $1 million in debt financing from Silicon Valley ... up automation and to advance its drug development efforts, ... new facility. "SVB has been an incredible ... the services a traditional bank would provide," said Dr. ...
(Date:6/23/2016)... ... , ... In a new case report published today in STEM CELLS Translational ... lymphedema after being treated for breast cancer benefitted from an injection of stem cells ... this debilitating, frequent side effect of cancer treatment. , Lymphedema refers to ...
Breaking Biology Technology:
(Date:3/29/2016)... RATON, Florida , March 29, 2016 ... or the "Company") LegacyXChange "LEGX" and SelectaDNA/CSI Protect are ... DNA in ink used in a variety of writing ... theft. Buyers of originally created collectibles from athletes on ... through forensic analysis of the DNA. ...
(Date:3/22/2016)... March 22, 2016 ... Sensors Market for Consumer Industry by Type (Image, ... Application (Communication & IT, Entertainment, Home Appliances, ... Forecast to 2022", published by MarketsandMarkets, the ... to reach USD 26.76 Billion by 2022, ...
(Date:3/21/2016)... 22, 2016 Unique technology ... for superior security   Xura, ... of secure digital communications services, today announced it is ... offer enterprise customers, particularly those in the Financial Services ... voice authentication within a mobile app, alongside, and in ...
Breaking Biology News(10 mins):