Genomic organization, cDNA sequence, bacterial expression, and purification of human seryl-tRNA synthase.
Genomic organization, cDNA sequence, bacterial expression, and purification of human seryl-tRNA synthase.
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基因组组织、cDNA 序列、细菌表达和人丝氨酰-tRNA 合酶的纯化。
DOI：
10.1111/j.1432-1033.1997.00077.x
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
M. Härtlein
中科院分区：
文献类型：
--
作者：
C. Vincent;N. Tarbouriech;M. Härtlein
本文报道了人胞浆丝氨酰-tRNA合成酶的cDNA序列和推导的一级序列，以及在大肠杆菌中的表达。对两个不同来源的人脑cDNA克隆进行测序，其中含有编码人丝氨酰-tRNA合成酶的重叠片段：HFBDN 14（胎脑克隆）和IB 48（婴儿脑克隆）。对于两个克隆，cDNA的5'区缺失。该5'区通过PCR方法使用人脑5' RACE-Ready cDNA文库获得。完整的cDNA序列使我们能够定义引物，分离和表征的内含子/外显子结构的Sers基因，由10个内含子和11个外显子。内含子的大小从283 bp到3000 bp以上，外显子的大小从71 bp到222 bp。人类酶的基因结构的可用性可能有助于澄清II类氨酰-tRNA合成酶分子进化的某些方面。人丝氨酰-tRNA合成酶已在大肠杆菌中表达。大肠杆菌，纯化（95%纯度，由SDS/PAGE测定）和动力学参数已测定其底物tRNA。人丝氨酰-tRNA合成酶序列（514个氨基酸残基）与大肠杆菌的丝氨酰-tRNA合成酶序列具有显著的同源性。大肠杆菌（25%）、酿酒酵母（40%）、拟南芥（41%）和秀丽隐杆线虫（60%）。已发表的哺乳动物丝氨酰-tRNA合成酶的部分序列与人酶非常相似（小鼠和中国仓鼠丝氨酰-tRNA合成酶的同一性分别为94%和92%）。人丝氨酰-tRNA合成酶与其他几种I类和II类人氨酰-tRNA合成酶类似，与细菌对应物明显相关，与人酶中鉴定的额外C-末端结构域和N-末端插入无关。在功能研究中，该酶氨酰化小牛肝tRNA和原核E。coli tRNA。
In this paper, we report the cDNA sequence and deduced primary sequence for human cytosolic seryl-tRNA synthetase, and its expression in Escherichia coli. Two human brain cDNA clones of different origin, containing overlapping fragments coding for human seryl-tRNA synthetase were sequenced: HFBDN14 (fetal brain clone); and IB48 (infant brain clone). For both clones the 5' region of the cDNA was missing. This 5' region was obtained via PCR methods using a human brain 5' RACE-Ready cDNA library. The complete cDNA sequence allowed us to define primers to isolate and characterize the intron/exon structure of the serS gene, consisting of 10 introns and 11 exons. The introns' sizes range from 283 bp to more than 3000 bp and the size of the exons from 71 bp to 222 bp. The availability of the gene structure of the human enzyme could help to clarify some aspects of the molecular evolution of class-II aminoacyl-tRNA synthetases. The human seryl-tRNA synthetase has been expressed in E. coli, purified (95% pure as determined by SDS/PAGE) and kinetic parameters have been measured for its substrate tRNA. The human seryl-tRNA synthetase sequence (514 amino acid residues) shows significant sequence identity with seryl-tRNA synthetases from E. coli (25%), Saccharomyces cerevisiae (40%), Arabidopsis thaliana (41%) and Caenorhabditis elegans (60%). The partial sequences from published mammalian seryl-tRNA synthetases are very similar to the human enzyme (94% and 92% identity for mouse and Chinese hamster seryl-tRNA synthetase, respectively). Human seryl-tRNA synthetase, similar to several other class-I and class-II human aminoacyl-tRNA synthetases, is clearly related to its bacterial counterparts, independent of an additional C-terminal domain and a N-terminal insertion identified in the human enzyme. In functional studies, the enzyme aminoacylates calf liver tRNA and prokaryotic E. coli tRNA.
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DOI：
10.1073/pnas.91.16.7435
发表时间：
1994-08-02
影响因子：
11.1
作者：
SHIBA, K;SUZUKI, N;NODA, T
通讯作者：
NODA, T
人腺苷脱氨酶 cDNA 序列，包括编码区和一个小内含子。
影响因子：
14.9
作者：
Wiginton,DA;Adrian,GS;Hutton,JJ
通讯作者：
Hutton,JJ
影响因子：
14.9
作者：
CAVENER, DR;RAY, SC
通讯作者：
RAY, SC
哺乳动物丝氨酰-tRNA 合成酶在体内与 mRNA 结合，与延伸因子 1 α 有同源性。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Miseta,A;Woodley,CL;Greenberg,JR;Slobin,LI
通讯作者：
Slobin,LI