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Unproductive transcript from MOPC21 cell line

Frequent amplification of the unproductive rearranged kappa variable IGKV-J gene from MOPC21, is observed during PCR of V genes from hybridomas.

References and accession numbers for the sequence of the unproductive rearranged IGKV-J from MOPC21:

• Strohal, R. et al., Nucleic Acids Res., 15, 2771 (1987). PMID: 3104882
• Carroll, W. L. et al., Molecular Immunology, 25, 991-995 (1988). PMID: 3146025
• Accession numbers: X05184, K00888 (g), M35669.

This unproductive rearranged IGKV-J transcript is found in the MOPC21-derived fusion partners (mouse myeloma cells) such as NS-1, P3-X63-Ag8.653, SP2/0 and P3U1 (P3X63Ag8U.1).

H⁺ and L⁺ indicate the expression of the productive Ig heavy and light transcripts in MOPC1 and P3-X63-Ag8. H^- and L^- indicate the absence of expression of the endogenous productive transcripts in the MOPC1 derived fusion partners. However the unproductive rearranged kappa transcript is still found.

Different approaches have been proposed to eliminate the unproductive transcript:

• Ribozyme cleavage: Duan, L. and Pomerantz, R. J., Nucleic Acids Res., 22, 5433-5438 (1994). PMID: 7816635
• Antisense-directed RNase H digestion: Ostermeier, C. and Michel, H., Nucleic Acids Res, 24, 1979-1980 (1996). PMID: 8657583

For more information on PCR and cloning of murine V genes, see also:

• Dattamajumdar, A. K. et al., Immunogenetics, 43, 141-151 (1996). PMID: 850098
• Nicholls, P. J. et al., J.Immunol. Methods, 165, 81-91 (1993). PMID: 8409471
• Kettleborough, C. A. et al., Eur. J. Immunol., 23, 206-211 (1993). PMID: 8419173

More on MOPC21 and P3:

• MOPC21 is a plasmacytoma (mouse IgG1 myeloma) originated by Potter et al. [1]
• P3K (= MOPC21) a cell line derived from the MOPC21 tumor adapted to continuous culture by Horibata and Harris [2]
• MOPC21 Disulphide bridges of the IgG1 [3]
• MOPC21 Complete amino acid kappa sequence [4]
• MOPC21 IG nucleotide sequences and Fab 3D structure
• Accession numbers for the unproductive rearranged IGH V-D-J rearrangement: X58634 (X63Ag8.653), D50398 (P3U1). The rearrangement is IGHV2-6*02-IGHD5-5*01-IGHJ3*01. Differences between X58634 and D50398 are most probably due to primers. (M.-P. Lefranc, 09/04/2011)

IMGT References:

[1]	Potter, M., Apella, E. and Geisser, S., J. Mol. Biol., 14, 361-372 (1965). PMID: 4160338
[2]	Horibata, K. and Harris, A.W., Exp. Cell Res., 60, 61-77 (1970). PMID: 5439579
[3]	Svasti, J. and Milstein, C., Biochem. J., 126, 837-850 (1972). PMID: 5073237
[4]	Svasti, J. and Milstein, C., Biochem. J., 128, 427-444 (1972). PMID: 4638343

See also:

• Cochet, O. et al., Biotechniques, 822, 818-820 (1999). PMID: 10337468
• Débat, H. et al., FASEB J., 15, 815-822 (2001). PMID: 11259400
• Juste, M. et al., Anal. Biochem., 349, 159-161 (2006). PMID: 16325758

Created: 27/04/2004
Last updated: Wednesday, 29-Jun-2011 17:01:52 CEST
Authors: Amina Mrani Dassouli and Marie-Paule Lefranc
Editor: Chantal Ginestoux
Contact: Marie-Paule Lefranc

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For any other use please contact Marie-Paule Lefranc Marie-Paule.Lefranc@igh.cnrs.fr.

     

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