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.

  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



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:

(Date:11/27/2015)... United Kingdom , Nov. 27, 2015 /PRNewswire/--  Mallinckrodt ... biopharmaceutical company, announced today that it has closed the ... (CMDS) business to Guerbet (GBT- NYSE Euronext) in a ... operations encompassed four manufacturing facilities and a total of ... 75 in the St. Louis ...
(Date:11/26/2015)... 2015 ... Research Laboratories, a leading independent and ... has formed a strategic partnership with ... Health for joint work on clinical ... ) , --> ,     ...
(Date:11/25/2015)... Nov. 25, 2015  PharmAthene, Inc. (NYSE MKT: PIP) ... a stockholder rights plan (Rights Plan) in an effort ... carryforwards (NOLs) under Section 382 of the Internal Revenue ... PharmAthene,s use of its NOLs could be substantially ... defined in Section 382 of the Code. In general, ...
(Date:11/25/2015)... HOLLISTON, Mass. , Nov. 25, 2015 ... a biotechnology company developing bioengineered organ implants for life-threatening ... will present at the LD Micro "Main Event" investor ... PT. The presentation will be webcast live and posted ... also be available at the conference for one-on-one meetings ...
Breaking Biology Technology:
(Date:10/29/2015)... JOLLA, Calif. , Oct. 29, 2015  The ... a new report titled, "DNA Synthesis and Biosecurity: Lessons ... well the Department of Health and Human Services guidance ... issued in 2010. --> ... but it also has the potential to pose unique ...
(Date:10/29/2015)... 29, 2015 Today, LifeBEAM , ... with 2XU, a global leader in technical performance ... hat with advanced bio-sensing technology. The hat will ... monitor key biometrics to improve overall training performance. ... two companies will bring together the most advanced technology, ...
(Date:10/26/2015)... LAS VEGAS , Oct. 26, 2015 ... an innovator in modern authentication and a founding member ... launch of its latest version of the Nok Nok™ ... to use standards-based authentication that supports existing and emerging ... Suite is ideal for organizations deploying customer-facing applications that ...
Breaking Biology News(10 mins):