Isolation and molecular characterization of bovine T-cell receptor genes

牛 T 细胞受体基因的分离和分子表征

• 批准号: 02806055
• 负责人: ISHIGURO Naotaka
• 金额: $ 1.22万
• 依托单位: Obihiro University of Agriculture and Veterinary Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02806055/
• 关键词: Bovine; Lymphocyte; T cell; Receptor; Gene rearrangement; cDNA; Nucleotide sequence; 牛; 遺伝子; 再構成

The nucleotide sequences of bovine T-cell receptor (TCR ; alpha, beta, gamma, and delta chains) genes which from cDNA libraries constructed from bovine peripheral blood lymphocytes and thymic lymphocytes were determined.1. Of 30 positive cDNA clones for bovine TCR alpha chain, 20 contained complete and correct rearrangements, whereas 4 contained only one Constant (C) region, and 6 were nonfunctional because of incomplete rearrangement. The bovine Calpha gene is composed of 140 amino acid residues with striking similarity to the Ca region of human and mouse.2. Of 38 CDNA clones for TCR beta chain, 22 were rearranged and contained the functional variable (V) gene segments. These clones were tentatively divided into nine TCR-V gene families which correspond to the human TCR-V family. Two different Cbeta gene segments were found, and both C regions were composed of 178 amino acid residues.3. Of 9 cDNA clones for bovine TCR gamma chain isolated, only 4 were-rearranged and contained specific … More V, Joining (J), and C gene segments, but the remaining 5 contained J and C or only C gene segments without the V gene segment. Three kinds of Cgamma were found and designated as Cgamma1, Cgamma2 and Cgamma3. Compared with human and mouse Cgamma, bovine Cgamma sequences are much longer, with 27-55 amino acids corresponding to the hinge and connector regions.4. Of three TCR delta chain CDNA clones isolated, two TCR V and three J gene segments were found. The nucleotide and deduced amino acid sequences (155 amino residues) of bovine C delta gene segment are well conserved among species.5. During the screening for bovine TCR alpha chain gene in a bovine lymphocyte CDNA library using mouse Calpha-encoding DNA as a probe, we isolated 11 CDNA clones encoding lactate dehydrogenase (LDH) -A isozyme. These LDH-A cDNA clones coded for 332 amino acid residues which protein was identified by cellogel electrophoresis as LDH-A isozyme.6. Sporadic bovine leukosis tumor cell lines were established by transplantation to BALB/C nu/nu mice from three types(calf, skin and thymic)of bovine lymphosarcoma, and examined for the expression of TCR alpha, beta, gamma and delta genes. Mature or truncated TCR-beta, -gamma and -delta transcripts were detected in the tumor cells, but all of them were found to be nonfunctional in cDNA cloning analysis. In the cDNA library constructed from the aberrant transcripts of the TCR-gamma gene of the thymic type of lymphosarcoma, a novel C region of the TCR gamma chain (Cgamma4) was isolated. Less

测定了牛外周血淋巴细胞和胸腺淋巴细胞构建的cDNA文库中牛T细胞受体（TCR；α、β、γ和δ链）基因的核苷酸序列。 1．在30个牛TCRα链阳性cDNA克隆中，20个含有完整且正确的重排，而4个仅含有1个恒定区（C），6个因重排不完全而无功能。牛Cα基因由140个氨基酸残基组成，与人和小鼠的Ca区极为相似。 2．在 38 个 TCR β 链 cDNA 克隆中，有 22 个经过重排并含有功能性可变 (V) 基因片段。这些克隆初步分为九个TCR-V基因家族，与人类TCR-V家族相对应。发现两个不同的Cbeta基因片段，两个C区均由178个氨基酸残基组成。 3．在分离的 9 个牛 TCR γ 链 cDNA 克隆中，只有 4 个被重排并包含特定的 V、连接 (J) 和 C 基因片段，但其余 5 个包含 J 和 C 或仅包含 C 基因片段而不含 V 基因片段。发现了三种Cgamma，并将其命名为Cgamma1、Cgamma2和Cgamma3。与人和小鼠Cgamma相比，牛Cgamma序列更长，铰链区和连接器区对应27-55个氨基酸。 4．在分离的三个 TCR δ 链 cDNA 克隆中，发现了两个 TCR V 和三个 J 基因片段。牛C delta基因片段的核苷酸序列和推导的氨基酸序列(155个氨基酸)在物种间具有良好的保守性。 5．在使用编码小鼠Cα的DNA作为探针在牛淋巴细胞cDNA文库中筛选牛TCRα链基因的过程中，我们分离了11个编码乳酸脱氢酶(LDH)-A同工酶的cDNA克隆。这些LDH-A cDNA克隆编码332个氨基酸残基，经凝胶电泳鉴定为LDH-A同工酶。6．将三种类型（小牛、皮肤和胸腺）牛淋巴肉瘤移植到BALB/C nu/nu小鼠中建立散发性牛白血病肿瘤细胞系，并检测TCRα、β、γ和δ基因的表达。在肿瘤细胞中检测到成熟或截短的TCR-β、-γ和-δ转录物，但在cDNA克隆分析中发现它们都没有功能。在由胸腺型淋巴肉瘤TCR-γ基因的异常转录本构建的cDNA文库中，分离出了TCRγ链的新C区（Cgamma4）。较少的

项目成果

Atsuko Tanaka: "Sequence and diversity of bovine T-cell receptor beta-chain genes." Immunogenetics. 32. 263-271 (1990)

Atsuko Tanaka：“牛 T 细胞受体 β 链基因的序列和多样性。”

Naotaka Ishiguro: "Primary structure of bovine lactate dehydrogenease-A isozyme and its synthesis in Escherichia coli." Gene. 91. 281-285 (1990)

Naotaka Ishiguro：“牛乳酸脱氢酶 A 同工酶的一级结构及其在大肠杆菌中的合成。”

Naotaka Ishiguro: "Sequence Analysis of bovine T-cell receptor genes and diagnostic application for bovine T-cell Iymphosarcoma." Proc. Jpn. Soc. Anim. Biochem.24. 48-61 (1990)

Naotaka Ishiguro：“牛 T 细胞受体基因的序列分析和牛 T 细胞淋巴肉瘤的诊断应用。”

Naotaka Ishiguro,Astuko Tanaka,Morikazu Shinagawa: "Sequence analvsis of bovine Tーcell receptorαchain" Immunogenetics. 31. 57-60 (1990)

Naotaka Ishiguro、Astuko Tanaka、Morikazu Shinakawa：“牛 T 细胞受体 α 链的序列分析”免疫遗传学。 31. 57-60 (1990)

Naotaka Ishiguro: "Sequence analysis of bovine T-cell receptor alpha chain." Immunogenetics. 31. 57-60 (1990)

Naotaka Ishiguro：“牛 T 细胞受体 α 链的序列分析。”