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Cell-type specific and regulated expression of human γ1 immunoglobulin genes in transgenic mice
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74 CELL-TYPE SPECIFIC AND REGULATED EXPRESSION OF HUMAN 71 IMMUNOGLOBULIN GENES IN TRANSGENIC MICE. K. Yamamura, A. Kudo, T. Ebihara, Y. Kumahara and T. Watanabe. Department of Medicine and Geriatrics, Osaka University Medical School, Fukushimaku, Osaka, 553, and Department of Immunology, Saga Medical School, Nabeshima, Saga, 840-01, Japan. A rearranged human ~'i immunoglobulin gene (HIGI) was cloned in charon 4A phage from human plasma cell leukemia cell line, ARH-77. About 200 copies of HIGI genes were introduced into fertilized mouse eggs in order to analyze cell-type specific and stage-specific enpression of HIGI gene. Total of four trangesmic mice were obtained. The HIGI mRNA was detected only in spleen cells. The level of mRNA and human yl chain were increased to ten to fifty folds after treatment with lipopolysaccharide but not with concanavalin A suggesting that the B lymphocyte specific and regulated expression of HIGI genes. Secretion of human yl chain was also observed. Mouse endogenous immunoglobulin production was not affected. This work was supported by a Grant-inAid for Special Project Research from the ministry of Education, Science, and Culture(Project No. 59480023), Japan.
MOLECULAR CLONING OF A cDNA CODING FOR A MOUSE ZONA PELLUCIDA PROTEIN M. Ringuette, D. Sobieski, S. Chamow, and J. Dean. Lab Cellular and Developmental Biol, NIH, Bethesda, Md. 20205 The murine zona pellucida surrounds maturing oocytes and is composed of three proteins; ZP-I, ZP-2 and ZP-3. One of these, ZP-3, has been shown to have species-specific sperm binding activity (Bleil and Wassarman, Cell 20:873, 1980). Using anti-zona monoclonal antibodies, we have identified a cDNA clone (~GT192) coding for ZP-3 from a ~ g t ] 1 ovarian library. After subcloning in pUC8, pGT192 was found by Northern analysis to hybridize uniquely to a 1.7kb poly(A)+ mRNA isolated from ovarian tissue; no hybridization was detected to somatic tissue mRNAs. There is no apparent DNA sequence rearrangement between germline and somatic tissues. These data are consistent with the hypothesis that the zona pellucida proteins represent a germline specific, developmentally gene family. Genomic Southern blots revealed considerable interspecies sequence divergence among mouse, rat and human and no hybridization to dog, pig and calf. 75 TWO 6-CRYSTALLIN GENES IN THE CHICKEN ARE TRANSCRIBED DIFFERENTLY DURING LENS DEVELOPMENT. K.Yasuda, H.Kondoh, Y.Shimura and T.S.Okada Dept. of Biophysics, Faculty of Science, University of Kyoto, Kyoto 606, Japan. As a clue to understand the molecular mechanism of specific gene expression, we took a close examination of the organization and structure of 6-erystallin genes by recombinant DNA technology and studied their expression by microinjection and in vitro transcription assays. Two 6-crystallin genes linked tandemly are separated by 5 kb of DNA and arranged in 5'-gene i-5 kb-gene 2-3'. They are transcribed in the same orientation. The cloned gene 1 injected into mouse lens ceils is expressed efficiently but the gene 2 not efficiently. Both genes ,however, are transcribed equally in in vitro transcription system. We showed that the DNA sequences around the CAT box is essential for its high level expression in lens cells. This region is not homologous to that of the gene 2 and this could explain that the gene 2 is not expressed efficiently during lens development.
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74 CELL-TYPE SPECIFIC AND REGULATED EXPRESSION OF HUMAN 71 IMMUNOGLOBULIN GENES IN TRANSGENIC MICE. K. Yamamura, A. Kudo, T. Ebihara, Y. Kumahara and T. Watanabe. Department of Medicine and Geriatrics, Osaka University Medical School, Fukushimaku, Osaka, 553, and Department of Immunology, Saga Medical School, Nabeshima, Saga, 840-01, Japan. A rearranged human ~'i immunoglobulin gene (HIGI) was cloned in charon 4A phage from human plasma cell leukemia cell line, ARH-77. About 200 copies of HIGI genes were introduced into fertilized mouse eggs in order to analyze cell-type specific and stage-specific enpression of HIGI gene. Total of four trangesmic mice were obtained. The HIGI mRNA was detected only in spleen cells. The level of mRNA and human yl chain were increased to ten to fifty folds after treatment with lipopolysaccharide but not with concanavalin A suggesting that the B lymphocyte specific and regulated expression of HIGI genes. Secretion of human yl chain was also observed. Mouse endogenous immunoglobulin production was not affected. This work was supported by a Grant-inAid for Special Project Research from the ministry of Education, Science, and Culture(Project No. 59480023), Japan.
MOLECULAR CLONING OF A cDNA CODING FOR A MOUSE ZONA PELLUCIDA PROTEIN M. Ringuette, D. Sobieski, S. Chamow, and J. Dean. Lab Cellular and Developmental Biol, NIH, Bethesda, Md. 20205 The murine zona pellucida surrounds maturing oocytes and is composed of three proteins; ZP-I, ZP-2 and ZP-3. One of these, ZP-3, has been shown to have species-specific sperm binding activity (Bleil and Wassarman, Cell 20:873, 1980). Using anti-zona monoclonal antibodies, we have identified a cDNA clone (~GT192) coding for ZP-3 from a ~ g t ] 1 ovarian library. After subcloning in pUC8, pGT192 was found by Northern analysis to hybridize uniquely to a 1.7kb poly(A)+ mRNA isolated from ovarian tissue; no hybridization was detected to somatic tissue mRNAs. There is no apparent DNA sequence rearrangement between germline and somatic tissues. These data are consistent with the hypothesis that the zona pellucida proteins represent a germline specific, developmentally gene family. Genomic Southern blots revealed considerable interspecies sequence divergence among mouse, rat and human and no hybridization to dog, pig and calf. 75 TWO 6-CRYSTALLIN GENES IN THE CHICKEN ARE TRANSCRIBED DIFFERENTLY DURING LENS DEVELOPMENT. K.Yasuda, H.Kondoh, Y.Shimura and T.S.Okada Dept. of Biophysics, Faculty of Science, University of Kyoto, Kyoto 606, Japan. As a clue to understand the molecular mechanism of specific gene expression, we took a close examination of the organization and structure of 6-erystallin genes by recombinant DNA technology and studied their expression by microinjection and in vitro transcription assays. Two 6-crystallin genes linked tandemly are separated by 5 kb of DNA and arranged in 5'-gene i-5 kb-gene 2-3'. They are transcribed in the same orientation. The cloned gene 1 injected into mouse lens ceils is expressed efficiently but the gene 2 not efficiently. Both genes ,however, are transcribed equally in in vitro transcription system. We showed that the DNA sequences around the CAT box is essential for its high level expression in lens cells. This region is not homologous to that of the gene 2 and this could explain that the gene 2 is not expressed efficiently during lens development.
32S