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

牛T细胞受体（TCR； Alpha，beta，Gamma和Delta链）基因的核苷酸序列，这些基因是从牛外周血血液淋巴细胞和胸腺淋巴细胞中构建的cDNA文库中的。1。在牛TCRα链的30个正cDNA克隆中，有20个包含完整和正确的重排，而4个仅包含一个常数（c）区域，而有6个由于不完整的重排而进行了无功能。牛Calpha基因由与人和小鼠CA区域相似的140个氨基酸残基组成。2。在38个用于TCRβ链的cDNA克隆中，有22个被重新排列，并包含功能变量（V）基因段。这些克隆分为九个与人类TCR-V家族相对应的TCR-V基因家族。发现了两个不同的CBETA基因段，两个C区域均由178个氨基酸残基组成3。分离的牛TCR伽马链的9个cDNA克隆中，只有4个重新编织并包含特定的……更多的V，加入（J）和C基因片段，但其余5个包含J和C基因段或仅C基因段，没有V基因段。发现并指定为Cgamma1，cgamma2和cgamma3。与人和小鼠Cgamma相比，牛Cgamma序列更长，其中27-55个氨基酸对应于铰链和连接器区域。4。在分离的三个TCR三角链cDNA克隆中，发现了两个TCR V和三个J基因段。在物种中，核苷酸和推导的氨基酸序列（155个氨基残留物）在物种之间得到很好的保守。5。在使用小鼠calpha编码DNA作为探针的牛淋巴细胞cDNA库中筛选牛TCRα链基因的过程中，我们分离了编码乳酸脱氢酶（LDH）-a同酶的11个cDNA克隆。这些LDH-A cDNA克隆编码为332个氨基酸残留物，通过大提琴电泳鉴定为LDH-A同工酶6。通过从牛淋巴瘤的三种类型（小牛，皮肤和胸腺）移植向BALB/C NU/NU小鼠，建立了零星的牛白细胞病细胞系，并检查了TCR Alpha，Beta，Beta，Gamma和Delta基因的表达。在肿瘤细胞中检测到成熟或截短的TCR -β，-gamma和-delta转录本，但在cDNA克隆分析中发现所有这些都不功能。在胸腺淋巴瘤类型的TCR-GAMMA基因的异常转录本构建的cDNA文库中，分离了TCR伽马链（CGAMMA4）的新型C区域。较少的

项目成果

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。”

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,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 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: "Primary structure of bovine lactate dehydrogenaseーAisozyme and its synthesis in Escherichia coli" Gene. 91. 281-285 (1990)

Naotaka Ishiguro：“牛乳酸脱氢酶-Aisozyme 的初级结构及其在大肠杆菌中的合成”基因。91. 281-285 (1990)