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Cochlear disorder associated with melanocyte anomaly in mice with a transgenic insertional mutation
6-O
1 COCHLEAR DISORDER ASSOCIATED WITH MELANOCYTE ANOMALY
INSERTIONAL
MUTATION.
MASAYOSHI
TACHIBANA.
IN MICE WITH A TRANSGENIC
Laborarorv of Molecular Bioloev. National Institute on Dcafncss
Imd
(Co-rexachers; Y.Hars*,D.Vyas*, CHodgkinson”, Neurological Disorders and Stroke and **NIDCD,
6-02
J.Fex**, K.Grundfxt**
and H.Arnheiter*.
*National
InstltuIe of
NIH, Bethesda, Maryland 20892, USA)
MYELIN BASIC PROTEIN GENE-RELATED PROTEIN (MGRPX MITSUKO SUGANO. TATSUNORI SEKI'1m
WI NAKAE, Advanced Research Laboratorv. Research and Development Center, _TOSHIBA corooratlon. 1 Komukai-Toshiba-cho. Salwalmku. Kawasaki 210. Jaoan and 'Deoartment of Anatomv, Juntendo ~_~ ~~__ ___- Universltv School of Medlclne.2-l-l Honno. Bunkvoumku. Tokyo 113. Japan transcrlpt is known to participate In the regulation of the gene expresslon thr.ough of double stranded RNA. Concerning the translated product from the antlsense transcript.however, it is not fully investigated even whether the translation occurs endogenously. the
AntIsense formation
We report the probability that a 30 kDa protein 1s translated from the sequence containing the antlsense transcript of myelin basic protein (MBP)gene. Thepartlal cDNA of the protein was Isolated. with a specific monoclonal antlbody. from the rat brain cDNA library (Clontech). Its nucleotide sequence was almost identical with the complementary sequence of the partial MBP mRNA corresponding to exon 4. 5 and 7. Then we have designated the protein as MBP gene-related protein (MGRP).MGRP was compared to MBP by lmmunoblotting and immunocytochemical analyses. Immunoblotting revealed that the ant1 MGRP monoclonal antibody did not recognize every MBP isoform. and lmmunocytochemical analyses revealed that MGRP mainly localized at the molecular layer, where MBP does not localize.in adult rat cerebellum. From these results,MGRP was regarded as not MBP but a distinct 30 kDa protein translated from the complementary sequence of MBP gene.
6-03
EXPRESSION OF THE POKKURI PROTEIN, A POSSIBLE TRANSCRIPTIONAL REGULATOR INVOLVED IN THE PHOTORECEPTOR FATE DETERMINATION IN DROSOPHILA. ITSUKO NIHONMATSU, KANAKO HIRATA, and DAISUKE YAMAMOTO. Mitsubishi Kasei Institute of Life Sciences, 11 Minamioova, Machida-shi. Tokvo JaRan. The Drosophila compound eye is composed of about 800 ommatidia, each of which contains eight identifiable photoreceptors, Rl-R8. We have isolated a gene pokkuri (pok), which plays a role in preventing overproduction of the R7 cell. Analysis of cDNA clones for pok has revealed that the gene encodes a .novel Ets related protein, which may function as a transcription factor. A pok cDNA containing the whole coding region was inserted in frame into an expression vector, pAR3251, driven by the bacteriophage T7 promoter. The construct was transformed into Escherichia coli BL21 cells, and it expressed a protein of approx. 90 kD which was absent in cells transformed with BL21 alone. We generated a monoclonal antibody, MAb nh1. using the bacterially expressed Pok protein as an antigen, and confirmed that the antibody recognized a protein of approx. 90 kD in the nuclear fruction of embryonic extract. Cellular and subcellular localizations of Pok protein in the developing eye are to be determined by immunohistochemistry with the antibody probe.
1 COCHLEAR DISORDER ASSOCIATED WITH MELANOCYTE ANOMALY
INSERTIONAL
MUTATION.
MASAYOSHI
TACHIBANA.
IN MICE WITH A TRANSGENIC
Laborarorv of Molecular Bioloev. National Institute on Dcafncss
Imd
(Co-rexachers; Y.Hars*,D.Vyas*, CHodgkinson”, Neurological Disorders and Stroke and **NIDCD,
6-02
J.Fex**, K.Grundfxt**
and H.Arnheiter*.
*National
InstltuIe of
NIH, Bethesda, Maryland 20892, USA)
MYELIN BASIC PROTEIN GENE-RELATED PROTEIN (MGRPX MITSUKO SUGANO. TATSUNORI SEKI'1m
WI NAKAE, Advanced Research Laboratorv. Research and Development Center, _TOSHIBA corooratlon. 1 Komukai-Toshiba-cho. Salwalmku. Kawasaki 210. Jaoan and 'Deoartment of Anatomv, Juntendo ~_~ ~~__ ___- Universltv School of Medlclne.2-l-l Honno. Bunkvoumku. Tokyo 113. Japan transcrlpt is known to participate In the regulation of the gene expresslon thr.ough of double stranded RNA. Concerning the translated product from the antlsense transcript.however, it is not fully investigated even whether the translation occurs endogenously. the
AntIsense formation
We report the probability that a 30 kDa protein 1s translated from the sequence containing the antlsense transcript of myelin basic protein (MBP)gene. Thepartlal cDNA of the protein was Isolated. with a specific monoclonal antlbody. from the rat brain cDNA library (Clontech). Its nucleotide sequence was almost identical with the complementary sequence of the partial MBP mRNA corresponding to exon 4. 5 and 7. Then we have designated the protein as MBP gene-related protein (MGRP).MGRP was compared to MBP by lmmunoblotting and immunocytochemical analyses. Immunoblotting revealed that the ant1 MGRP monoclonal antibody did not recognize every MBP isoform. and lmmunocytochemical analyses revealed that MGRP mainly localized at the molecular layer, where MBP does not localize.in adult rat cerebellum. From these results,MGRP was regarded as not MBP but a distinct 30 kDa protein translated from the complementary sequence of MBP gene.
6-03
EXPRESSION OF THE POKKURI PROTEIN, A POSSIBLE TRANSCRIPTIONAL REGULATOR INVOLVED IN THE PHOTORECEPTOR FATE DETERMINATION IN DROSOPHILA. ITSUKO NIHONMATSU, KANAKO HIRATA, and DAISUKE YAMAMOTO. Mitsubishi Kasei Institute of Life Sciences, 11 Minamioova, Machida-shi. Tokvo JaRan. The Drosophila compound eye is composed of about 800 ommatidia, each of which contains eight identifiable photoreceptors, Rl-R8. We have isolated a gene pokkuri (pok), which plays a role in preventing overproduction of the R7 cell. Analysis of cDNA clones for pok has revealed that the gene encodes a .novel Ets related protein, which may function as a transcription factor. A pok cDNA containing the whole coding region was inserted in frame into an expression vector, pAR3251, driven by the bacteriophage T7 promoter. The construct was transformed into Escherichia coli BL21 cells, and it expressed a protein of approx. 90 kD which was absent in cells transformed with BL21 alone. We generated a monoclonal antibody, MAb nh1. using the bacterially expressed Pok protein as an antigen, and confirmed that the antibody recognized a protein of approx. 90 kD in the nuclear fruction of embryonic extract. Cellular and subcellular localizations of Pok protein in the developing eye are to be determined by immunohistochemistry with the antibody probe.