Somatic mutation of immunoglobulin V genes in vitro

Science

Volumn 271, Issue 5253, 1996, Pages 1285-1289

  Källberg, Eva ^a   Jainandunsing, Sandra ^b   Gray, David ^b   Leanderson, Tomas ^a  

^a LUND UNIVERSITY   (Sweden)

^b HAMMERSMITH HOSPITAL   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

CD40 ANTIGEN; LIPOPOLYSACCHARIDE; LYMPHOCYTE MEMBRANE IMMUNOGLOBULIN;

ANIMAL CELL; ARTICLE; B LYMPHOCYTE; CONTROLLED STUDY; CROSS LINKING; IMMUNOGLOBULIN GENE; MOUSE; NONHUMAN; POINT MUTATION; PRIORITY JOURNAL; SOMATIC MUTATION; T LYMPHOCYTE;

ANIMALIA;

EID: 0029879794     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.271.5253.1285     Document Type: Article

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24. κ5-specific primer (5′-ACGTTTCAGCTCCAGCTTGGTC-CCA-3′) DNA sequencing was performed with T7 DMA polymerase or with Taq polymerase with the use of dior dye-deoxy terminators
25. The source of CD40 ligand was the transfectant cell line K47 (G. Wohlleben et al., Int Immunol., in press) that constitutively expresses CD40 ligand (C040L)
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27. κOx1 rearrangements were amplified by RT-PCR as described above and cloned into pGem3Z vector. Random plasmids containing inserts were selected and sequenced. The measured Taq polymerase error under these conditions is less than one mutation per 1000 bases sequenced.
28. Supported by grants from the Swedish Medical Research Council, the Alfred Östetlund Foundation, the Crafoord Foundation, the Swedish Cancer Society, the British Medical Research Council, Kocks Foundation, and the Wellcome Trust