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. 2002 Jan 7;195(1):143–149. doi: 10.1084/jem.20011524

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Copyright © 2002, The Rockefeller University Press

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Figure 1. — Disruption of the Bam32 gene in chicken DT40 B cells. (A) Partial restriction map of chicken Bam32 locus and expected structure of the targeted Bam32 alleles. The restriction endonuclease cleavage sites are abbreviated: X, XbaI; E, EcoRI. (B) Southern analysis of genomic DNA from wild-type DT40 cells (lane 1), neo-targeted cells (+/−, lane 2), and neo- and hisD-targeted cells (−/−, lane 3 and 4). Xba I-digested genomic DNA separated on an agarose gel was blotted and hybridized with a chicken Bam32 DNA probe shown in A. Wild-type (WT) and targeted alleles show 15.5- or 5.0-kb fragments, respectively. (C) Northern analysis using a chicken cDNA probe for Bam32 (top) and β-actin (bottom). Wild-type (WT) DT40 cells contain a hybridizable RNA species of ∼3,000 nucleotides. (D) Cell surface expression of BCR. BCR expression on the surface of wild-type (WT) and two independent Bam32-deficient (M80–11, M82–5) DT40 cells was analyzed by flow cytometry. Unstained cells were used as negative controls.

Figure 1. — Disruption of the Bam32 gene in chicken DT40 B cells. (A) Partial restriction map of chicken Bam32 locus and expected structure of the targeted Bam32 alleles. The restriction endonuclease cleavage sites are abbreviated: X, XbaI; E, EcoRI. (B) Southern analysis of genomic DNA from wild-type DT40 cells (lane 1), neo-targeted cells (+/−, lane 2), and neo- and hisD-targeted cells (−/−, lane 3 and 4). Xba I-digested genomic DNA separated on an agarose gel was blotted and hybridized with a chicken Bam32 DNA probe shown in A. Wild-type (WT) and targeted alleles show 15.5- or 5.0-kb fragments, respectively. (C) Northern analysis using a chicken cDNA probe for Bam32 (top) and β-actin (bottom). Wild-type (WT) DT40 cells contain a hybridizable RNA species of ∼3,000 nucleotides. (D) Cell surface expression of BCR. BCR expression on the surface of wild-type (WT) and two independent Bam32-deficient (M80–11, M82–5) DT40 cells was analyzed by flow cytometry. Unstained cells were used as negative controls.