Organization and structure of NADH-dependent glutamate synthase gene from rice plants

Satoshi Goto; Takumi Akagawa; Soichi Kojima; Toshihiko Hayakawa; Tomoyuki Yamaya

doi:10.1016/S0167-4838(98)00142-3

Organization and structure of NADH-dependent glutamate synthase gene from rice plants

Satoshi Goto, Takumi Akagawa, Soichi Kojima, Toshihiko Hayakawa, Tomoyuki Yamaya

AGR - Agricultural Chemistry

Tohoku University

Research output: Contribution to journal › Article › peer-review

48 Citations (Scopus)

Abstract

Genomic clones for NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) were obtained from a genomic library of rice (Oryza sativa L. cv. Sasanishki). A genomic clone (λOS42, 14 kb) covered an entire structural gene and a 3.7 kb 5'-upstream region from the first methionine. Another clone (λOS23, 14 kb) contained a 2.8 kb 3'-downstream region from the stop codon. A 7047 bp long clone (λOSR51) consisting of full length cDNA for NADH-GOGAT was isolated from a cDNA library prepared using mRNA from roots of rice seedlings treated with 1 mM NH₄Cl for 12 h. The presumed transcribed region (11.7 kb) consisted of 23 exons separated by 22 introns. Rice NADH-GOGAT is synthesized as a 2166 amino acid protein with a molecular mass of 236.7 kDa that includes a 99 amino acid presequence. DNA gel blot analysis suggested that NADH-GOGAT occurred as a single gene in rice. Primer extension experiments map the transcription start of NADH-GOGAT to identical positions. The 3.7 kb 5'-upstream region was able to transiently express a reporter gene in cultured rice cells. Putative motifs related to the regulation of NADH-GOGAT gene expression were looked for within the 5'-upstream region by database. Copyright (C) 1998 Elsevier Science B.V.

Original language	English
Pages (from-to)	298-308
Number of pages	11
Journal	Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume	1387
Issue number	1-2
DOIs	https://doi.org/10.1016/S0167-4838(98)00142-3
Publication status	Published - 1998 Sept 8

Keywords

cDNA
Gene structure
NADH-glutamate synthase
Rice, Oryza sativa

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10.1016/S0167-4838(98)00142-3

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@article{3bef08358fec402298556f885ba2640c,

title = "Organization and structure of NADH-dependent glutamate synthase gene from rice plants",

abstract = "Genomic clones for NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) were obtained from a genomic library of rice (Oryza sativa L. cv. Sasanishki). A genomic clone (λOS42, 14 kb) covered an entire structural gene and a 3.7 kb 5'-upstream region from the first methionine. Another clone (λOS23, 14 kb) contained a 2.8 kb 3'-downstream region from the stop codon. A 7047 bp long clone (λOSR51) consisting of full length cDNA for NADH-GOGAT was isolated from a cDNA library prepared using mRNA from roots of rice seedlings treated with 1 mM NH4Cl for 12 h. The presumed transcribed region (11.7 kb) consisted of 23 exons separated by 22 introns. Rice NADH-GOGAT is synthesized as a 2166 amino acid protein with a molecular mass of 236.7 kDa that includes a 99 amino acid presequence. DNA gel blot analysis suggested that NADH-GOGAT occurred as a single gene in rice. Primer extension experiments map the transcription start of NADH-GOGAT to identical positions. The 3.7 kb 5'-upstream region was able to transiently express a reporter gene in cultured rice cells. Putative motifs related to the regulation of NADH-GOGAT gene expression were looked for within the 5'-upstream region by database. Copyright (C) 1998 Elsevier Science B.V.",

keywords = "cDNA, Gene structure, NADH-glutamate synthase, Rice, Oryza sativa",

author = "Satoshi Goto and Takumi Akagawa and Soichi Kojima and Toshihiko Hayakawa and Tomoyuki Yamaya",

note = "Funding Information: We thank Dr. Carroll Vance, University of Minnesota, USA, for his generous gift of alfalfa nodule NADH-GOGAT cDNA. We would also like to thank Dr. Ann Oaks, University of Guelph, Canada and Dr. Laura Hopkins, University of St. Andrews, UK, for their helpful comments and critical reading of the manuscript. Technical supports by N. Hirose, N. Sakurai, S. Watanabe, K. Azuma, Y. Shimura, A. Sakai, and M. Hino in our lab are greatly acknowledged. This work was supported in part by a program of Research for the Future from the Japan Society for the Promotion of Science (JSPS-RFTF96L00604), in part by Grant-in-Aids for Scientific Research on Priority Area (Nos. 09274101 and 09274102), and in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan (No. 08044187).",

year = "1998",

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doi = "10.1016/S0167-4838(98)00142-3",

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T1 - Organization and structure of NADH-dependent glutamate synthase gene from rice plants

AU - Goto, Satoshi

AU - Akagawa, Takumi

AU - Kojima, Soichi

AU - Hayakawa, Toshihiko

AU - Yamaya, Tomoyuki

N1 - Funding Information: We thank Dr. Carroll Vance, University of Minnesota, USA, for his generous gift of alfalfa nodule NADH-GOGAT cDNA. We would also like to thank Dr. Ann Oaks, University of Guelph, Canada and Dr. Laura Hopkins, University of St. Andrews, UK, for their helpful comments and critical reading of the manuscript. Technical supports by N. Hirose, N. Sakurai, S. Watanabe, K. Azuma, Y. Shimura, A. Sakai, and M. Hino in our lab are greatly acknowledged. This work was supported in part by a program of Research for the Future from the Japan Society for the Promotion of Science (JSPS-RFTF96L00604), in part by Grant-in-Aids for Scientific Research on Priority Area (Nos. 09274101 and 09274102), and in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan (No. 08044187).

PY - 1998/9/8

Y1 - 1998/9/8

N2 - Genomic clones for NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) were obtained from a genomic library of rice (Oryza sativa L. cv. Sasanishki). A genomic clone (λOS42, 14 kb) covered an entire structural gene and a 3.7 kb 5'-upstream region from the first methionine. Another clone (λOS23, 14 kb) contained a 2.8 kb 3'-downstream region from the stop codon. A 7047 bp long clone (λOSR51) consisting of full length cDNA for NADH-GOGAT was isolated from a cDNA library prepared using mRNA from roots of rice seedlings treated with 1 mM NH4Cl for 12 h. The presumed transcribed region (11.7 kb) consisted of 23 exons separated by 22 introns. Rice NADH-GOGAT is synthesized as a 2166 amino acid protein with a molecular mass of 236.7 kDa that includes a 99 amino acid presequence. DNA gel blot analysis suggested that NADH-GOGAT occurred as a single gene in rice. Primer extension experiments map the transcription start of NADH-GOGAT to identical positions. The 3.7 kb 5'-upstream region was able to transiently express a reporter gene in cultured rice cells. Putative motifs related to the regulation of NADH-GOGAT gene expression were looked for within the 5'-upstream region by database. Copyright (C) 1998 Elsevier Science B.V.

AB - Genomic clones for NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) were obtained from a genomic library of rice (Oryza sativa L. cv. Sasanishki). A genomic clone (λOS42, 14 kb) covered an entire structural gene and a 3.7 kb 5'-upstream region from the first methionine. Another clone (λOS23, 14 kb) contained a 2.8 kb 3'-downstream region from the stop codon. A 7047 bp long clone (λOSR51) consisting of full length cDNA for NADH-GOGAT was isolated from a cDNA library prepared using mRNA from roots of rice seedlings treated with 1 mM NH4Cl for 12 h. The presumed transcribed region (11.7 kb) consisted of 23 exons separated by 22 introns. Rice NADH-GOGAT is synthesized as a 2166 amino acid protein with a molecular mass of 236.7 kDa that includes a 99 amino acid presequence. DNA gel blot analysis suggested that NADH-GOGAT occurred as a single gene in rice. Primer extension experiments map the transcription start of NADH-GOGAT to identical positions. The 3.7 kb 5'-upstream region was able to transiently express a reporter gene in cultured rice cells. Putative motifs related to the regulation of NADH-GOGAT gene expression were looked for within the 5'-upstream region by database. Copyright (C) 1998 Elsevier Science B.V.

KW - cDNA

KW - Gene structure

KW - NADH-glutamate synthase

KW - Rice, Oryza sativa

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Organization and structure of NADH-dependent glutamate synthase gene from rice plants

Abstract

Keywords

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