Expression Cloning of Rat cDNA Encoding UDP-galactose:GD2 β1,3-galactosyltransferase That Determines the Expression of GD1b/GM1/GA1 *
Expression Cloning of Rat cDNA Encoding UDP-galactose:GD2 β1,3-galactosyltransferase That Determines the Expression of GD1b/GM1/GA1 *
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编码 UDP-半乳糖的大鼠 cDNA 的表达克隆：决定 GD1b/GM1/GA1 表达的 GD2 β1,3-半乳糖基转移酶 *
DOI：
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发表时间：
1997
影响因子：
4.8
通讯作者：
Kazuro Furukawa
中科院分区：
文献类型：
--
作者：
H. Miyazaki;S. Fukumoto;M. Okada;T. Hasegawa;Keiko Furukawa;Kazuro Furukawa
使用抗GD1b单克隆抗体，进行β1,3-半乳糖基转移酶基因(EC 2.4.1.62)的cDNA的表达克隆。 KF4C、转染多瘤T抗原基因的小鼠黑色素瘤B16和GM2/GD2合酶cDNA用作cDNA文库转染的受体细胞系。 GD3合酶的cDNA克隆pD3T-31与使用pcDNAI表达载体从大鼠脑RNA制备的cDNA文库共转染。分离的cDNA克隆pM1T-9预测具有4个氨基酸的胞质结构域、21个氨基酸的跨膜结构域和具有346个氨基酸的大催化结构域的II型膜蛋白。将 cDNA 克隆引入先前转染 GM2/GD2 合酶基因的小鼠黑色素瘤系 B16 中，导致 GM1 的新合成。用 pM1T-9 和 GD3 合酶 cDNA 共转染细胞系导致 GD1b 和 GM1 的表达。此外，将pM1T-9导入预先转染GM2/GD2合酶基因的L细胞（缺乏GM3合酶）中，导致asialo-GM1的明确表达。这些结果表明 GD1b/GM1/GA1 合酶是相同的，正如之前基于酶学分析所建议的那样。在 β1,3-半乳糖基转移酶基因与来自不同大鼠组织的总 RNA 的 Northern 印迹中，1.6 KB 的 mRNA 在脾、胸腺、肾和睾丸中强烈表达。然而，该基因在成人脑组织中的表达水平并不是特别高。另一方面，该基因在胚胎第12天的大鼠大脑中高水平表达，并在出生前后达到峰值，然后在成年大脑中降至低水平。
Using an anti-GD1b monoclonal antibody, expression cloning of a cDNA for the β1,3-galactosyltransferase gene (EC 2.4.1.62) was performed. KF4C, mouse melanoma B16 transfected with polyoma T antigen gene, and GM2/GD2 synthase cDNA was used as a recipient cell line for the cDNA library transfection. A cDNA clone of GD3 synthase, pD3T-31 was co-transfected with a cDNA library prepared from rat brain RNA using the pcDNAI expression vector. The isolated cDNA clone pM1T-9 predicted a type II membrane protein with 4 amino acids of cytoplasmic domain, 21 amino acids of transmembrane region, and a large catalytic domain with 346 amino acids. Introduction of the cDNA clone into a mouse melanoma line B16 previously transfected with a GM2/GD2synthase gene resulted in the neo-synthesis of GM1. Co-transfection of the cell line with pM1T-9 and a GD3synthase cDNA resulted in the expression of GD1b as well as GM1. Moreover, introduction of pM1T-9 into L cell (lacking GM3 synthase), previously transfected with GM2/GD2 synthase gene, resulted in the definite expression of asialo-GM1. These results indicated that GD1b/GM1/GA1 synthases were identical, as previously suggested based on enzymological analysis. In Northern blots of the β1,3-galactosyltransferase gene with total RNA from various rat tissues, a 1.6-kilobase mRNA was strongly expressed in spleen, thymus, kidney, and testis. However, the expression level of the gene in the adult brain tissue was not especially high. On the other hand, this gene was expressed at high levels in the rat brain of embryonal day 12, and reached a peak at around birth, then fell to low level in the adult brain.
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DOI：
10.1016/s0021-9258(19)40170-1
发表时间：
1990-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
F. Yamamoto;J. Marken;T. Tsuji;T. White;H. Clausen;S. Hakomori
通讯作者：
F. Yamamoto;J. Marken;T. Tsuji;T. White;H. Clausen;S. Hakomori
牛 α 1----3-半乳糖基转移酶：cDNA 克隆的分离和表征。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Joziasse,DH;Shaper,JH;VandenEijnden,DH;VanTunen,AJ;Shaper,NL
通讯作者：
Shaper,NL
DOI：
10.1073/pnas.91.2.728
发表时间：
1994-01-18
影响因子：
11.1
作者：
IOFFE, E;STANLEY, P
通讯作者：
STANLEY, P
DOI：
10.1016/s0021-9258(17)39801-0
发表时间：
1984-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
E. Bremer;S. Hakomori;D. Bowen-Pope;E. Raines;R. Ross
通讯作者：
E. Bremer;S. Hakomori;D. Bowen-Pope;E. Raines;R. Ross
黑色素瘤和神经母细胞瘤细胞中神经节苷脂生物合成的糖基化途径：相对糖基转移酶水平决定神经节苷脂模式。
影响因子：
11.2
作者：
Ruan,S;Lloyd,KO
通讯作者：
Lloyd,KO