Tsui F W, Siminovitch L
Nucleic Acids Res. 1987 Apr 24;15(8):3349-67. doi: 10.1093/nar/15.8.3349.
A full length cDNA clone that codes for human histidyl-tRNA synthetase (HRS) and cDNA clones that span the full length transcript of hamster HRS have been isolated. The full length human HRS cDNA was expressed after transfection into Cos 1 cells and a CHO ts mutant defective in the gene for HRS. The complete nucleotide sequence of the hamster and human gene were obtained and extensive homologies were observed in three regions on comparing these sequences between themselves and with the sequence of HRS derived from yeast. These results provide unequivocal evidence that we have indeed cloned the hamster and human gene for HRS. Three overlapping phage recombinants containing the complete hamster chromosomal gene for HRS have also been isolated. The genomic HRS is divided into 13 exons. The precise locations of each of the 5' and 3' exon-intron boundaries were defined by sequencing the appropriate regions of the cloned genomic DNA and aligning them with the sequence of HRS cDNAs. These studies provide the basis for future structural and functional analysis of the gene for HRS. In particular, it will be of interest to examine if different exons of HRS correlate to different domains of the HRS polypeptide.
编码人组氨酰 - tRNA合成酶（HRS）的全长cDNA克隆以及跨越仓鼠HRS全长转录本的cDNA克隆已被分离出来。将全长人HRS cDNA转染到Cos 1细胞和HRS基因有缺陷的CHO ts突变体中后，其得以表达。获得了仓鼠和人基因的完整核苷酸序列，在将它们彼此之间以及与源自酵母的HRS序列进行比较时，在三个区域观察到了广泛的同源性。这些结果提供了明确的证据，表明我们确实克隆了仓鼠和人的HRS基因。还分离出了三个包含完整仓鼠HRS染色体基因的重叠噬菌体重组体。基因组HRS被分为13个外显子。通过对克隆的基因组DNA的适当区域进行测序并将它们与HRS cDNA序列比对，确定了5'和3'外显子 - 内含子边界各自的精确位置。这些研究为未来对HRS基因进行结构和功能分析奠定了基础。特别是，研究HRS的不同外显子是否与HRS多肽的不同结构域相关将是很有意义的。
