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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 (IMGT/LIGM-DB).

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.

In NS0, a non-IG secreting H-L- cell line, derived from NS-1, a frameshift mutation in the L-KAPPA mRNA is responsible of the loss of the kappa chain expression. Hybridomas made with NS0 contain this unproductive transcript.

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
Restriction enzyme digestion step using BciVI: Juste M. et al., Anal. Biochem., 349, 159-161 (2006). PMID: 16325758

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
MOPC21 Complete sequencing of the L-KAPPA using the dideoxynucleotide method of RNA sequencing (V 00810) [5]
In that sequence, one nucleotide mutation leads to an amino acid change: N42 (codon aac), instead of K42 (codon aag) (IMGT unique numbering).
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
[5]	Hamlyn P.H. et al. Nucleic Acids Res. 9, 4485-4494 (1981). PMID: 6170937

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