The Expression Mechanism of Leghemoglobin Geno of Leguminous Plant.

豆科植物豆血红蛋白基因的表达机制。

• 批准号: 60540438
• 负责人: HIGASHI Shiro
• 金额: $ 1.02万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60540438/
• 关键词: Legume-root nodule; Leghemoglobin; Lb-synthesized cell; Lb-induction gene(Lb-ind gene) Lb-cDNA; Lb-cDNA; Lb-ind gene; transfarmants; 根粒菌; 感染機構; Lb-発現機構

Leghemoglobins (Lbs) are small monomeric heme proteins synthesized exclusively in the root nodules that develope due to the rhizobial infection to leguminous plant. The induction of Lb-genes occurs only after bacterial infection but is independent of the expression of nif-gene in bacteroid. The Lb-gene expression of root cells are induced by some signals derived from the invaded bacteria. The signals are not Known about the details although nod- and nif-gene have been analyzed about the loci on Sym-plasmid and nucleotide sequences.The cell, leghemoglobin is synthesized at the most early step of nodulation, has been determined by an indirect-immunological technique.For analysis the expression of Lb-induction gene(Lb-ind gene), Lb-gene has been cloned with the mRNA of root nodule : reverse transcriptase system. The total DNA from R. trifolii 4S(Nod^+, Lb^+, Fix^+) was isolated and digested with Sau3AI. The DNA fragments were ligated with cosmid vector pCKS 13 and packaged in <lambda>-phage head. The recombinant DNA including in E. coli cells were transferred to R. trifolii Qn1(Nod^+,Lb^-, Fix^-) by triparental mating method, and then inoculated to seedilings of white clover. A strain NL18(Nod^+, Lb^+, Fix^-) was screened out form a nodule. The locus and size of Lb-ind gene on the cloning DNA from R. trifolii 4S was discussed in this study.

豆血红蛋白（Lbs）是豆科植物根瘤菌侵染后产生的小单体血红素蛋白。Lb基因的诱导只发生在细菌感染后，但不依赖于类菌体中nif基因的表达。根细胞Lb基因的表达受来自入侵细菌的一些信号的诱导。虽然nod-和nif-基因在Sym-质粒上的位置和核苷酸序列已经分析过，但信号的细节还不清楚。通过间接免疫技术，已经确定了细胞，豆血红蛋白在形成的最早阶段合成。为了分析Lb-诱导基因（Lb-ind基因）的表达，Lb-基因已经与根瘤mRNA：逆转录酶系统一起克隆。从R. trifolii 4S（Nod^+，Lb^+，Fix^+）经Sau 3AI酶切后，用琼脂糖凝胶电泳（PAGE）鉴定。DNA片段与粘粒载体pCKS 13连接，包装在<lambda>噬菌体头部.将重组DNA插入E. coli细胞转入R. trifolii Qn 1（Nod^+，Lb^-，Fix^-），然后接种于白色三叶草幼苗。从根瘤菌中筛选出一株NL 18（Nod^+，Lb^+，Fix^-）。Lb-ind基因在克隆DNA上的位置和大小。对三叶草4S进行了研究。

项目成果

Shiro Higashi;Kazuya Kushiyama;Mikiko Abe.: Canadian Journal Microbiology. 32. 947-952 (1986)

Shiro Higashi；Kazuya Kushiyama；Mikiko Abe。：加拿大微生物学杂志。

東四郎: 微生物. 3. (1987)

俊郎：微生物3。（1987）

Shiro Higashi;Mikiko Abe;Gloria D.Reyes;Ireneo J.Manguiat.: Journal of General and Applied Microbiology.33. 241-245 (1987)

Shiro Higashi;Mikiko Abe;Gloria D.Reyes;Ireneo J.Manguiat.：普通与应用微生物学杂志.33。

Shiro Higashi, Mikiko Abe, Gloria D. Reyes and Ireneo J. Manguiat: "Electron microscopic studies of the root nodule of Pterocapus indicus." Journal of General and Applied Microbiology. 3. 346-355 (1987)

Shiro Higashi、Mikiko Abe、Gloria D. Reyes 和 Ireneo J. Manguiat：“Pterocapus indicus 根瘤的电子显微镜研究”。

Shiro Higashi;Kazumi Sakai;Toshiki Uchiumi;Mikiko Abe: Journal Bacteriology.(1987)

Shiro Higashi；Kazumi Sakai；Toshiki Uchiumi；Mikiko Abe：细菌学杂志。（1987）