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note
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Murine genomic Igβ DNA clones were isolated from a 129/Sv strain genomic library. The long arm of the targeting vector consisted of 10 kilobases (kb) of DMA 5′ to the Eco RV site, which is 1 kb upstream of the Igβ promoter The short arm was a 1.2-kb fragment from the Kpn I site in the second exon to the Xba I site in the third intron. Homologous insertion resulted in a deletion of the promoter, the first exon, and the portion of the second exon that is 5′ to the Kpn I site. The targeting construct was lineanzed with Not I and transfected into CJ7 (22) embryonic stem (ES) cells. Two hundred ES cell clones were screened, and four clones carrying the mutation in Igβ were identified by Southern blotting. All four were injected into C57BI/6 blastocysts, and three of the four clones produced chimeric mice that transmitted the mutation.
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Hster were as described previously (10, 16, 25)
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15844391003
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note
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Lymph nodes and spleens were photographed at magnifications of ×X10 and ×25, respectively. Tissues from 5-week-old mice were fixed with formalin, and tissue sections were stained with hematoxylin and eosin.
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note
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+ gale are shown in each panel. Analysis was on a FACstar-plus system with the Lysis-II program (Becton Dickinson).
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H1 and 2, respectively. Control primers were from the J-CH1 intervening sequence (IVS) (24).
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note
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H (17) and a 650-bp Hind III-Pst I fragment that covers lgμ-CH4
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note
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We thank M. Sanchez and T Dlohery for the FACS analyses; K. Sokol for histology; H. Petree, A Fienberg, X. Qin, and Z. Misulovin for technical advice; and S, Gezelter and F. Isdell for cell sorting.
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