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)... 23, 2016  Blueprint Bio, a company dedicated to ... medical community, has closed its Series A funding round, ... "We have received a commitment from Forentis ... need to meet our current goals," stated Matthew ... runway to complete validation on the current projects in ...
(Date:6/23/2016)... -- On Wednesday, June 22, 2016, the NASDAQ ... Dow Jones Industrial Average edged 0.27% lower to finish at ... Stock-Callers.com has initiated coverage on the following equities: Infinity Pharmaceuticals ... NKTR ), Aralez Pharmaceuticals Inc. (NASDAQ: ARLZ ... Learn more about these stocks by accessing their free trade ...
(Date:6/23/2016)... ... 23, 2016 , ... Regulatory Compliance Associates® Inc. (RCA), a ... webinar on Performing Quality Investigations: Getting to Root Cause. This ... charge. , Incomplete investigations are still a major concern to the Regulatory Authorities ...
(Date:6/23/2016)... June 22, 2016  Amgen (NASDAQ: AMGN ... QB3@953 life sciences incubator to accelerate the ... shared laboratory space at QB3@953 was created to help ... obstacle for many early stage organizations - access to ... sponsorship, Amgen launched two "Amgen Golden Ticket" awards, providing ...
Breaking Biology Technology:
(Date:4/28/2016)... April 28, 2016 First quarter 2016:   ... 966% compared with the first quarter of 2015 The ... 589.1 M (loss: 18.8) and the operating margin was 40% (-13) ... Cash flow from operations was SEK 249.9 M (21.2) ... guidance is unchanged, SEK 7,000-8,500 M. The operating margin ...
(Date:4/15/2016)... , April 15, 2016 ... the,  "Global Gait Biometrics Market 2016-2020,"  report to ... http://photos.prnewswire.com/prnh/20160330/349511LOGO ) , ,The global gait biometrics ... of 13.98% during the period 2016-2020. ... angles, which can be used to compute factors ...
(Date:3/29/2016)... March 29, 2016 LegacyXChange, Inc. ... "LEGX" and SelectaDNA/CSI Protect are pleased to announce our ... in a variety of writing instruments, ensuring athletes signatures ... created collectibles from athletes on LegacyXChange will be assured ... the DNA. Bill Bollander , CEO ...
Breaking Biology News(10 mins):