McNally M T, Free S J
Department of Biological Sciences, State University of New York, Buffalo 14260.
Curr Genet. 1988 Dec;14(6):545-51. doi: 10.1007/BF00434079.
Using differential hybridization, the cDNA copy of a Neurospora gene coding for an abundant glucose-repressible mRNA (grg-1) has been isolated. The cDNA was used to clone the genomic copy, and both were sequenced. The cDNA is nearly full length and contains putative translational start and termination codons. Conceptual translation indicates that grg-1 mRNA could direct the synthesis of a 7,000 molecular weight polypeptide. The genomic clone, contained in an 1,888 bp PvuII fragment, encompasses the entire cDNA as well as 838 bp of 5' and 369 bp of 3' flanking sequence. Comparison of the cDNA and genomic clones revealed the presence of two short introns in potential protein-coding sequences. grg-1 message levels were found to increase within minutes following the onset of glucose deprivation and rise 50 fold during the first 90 min of derepression.
利用差异杂交技术,已分离出编码一种丰富的葡萄糖可阻遏mRNA(grg-1)的粗糙脉孢菌基因的cDNA拷贝。该cDNA被用于克隆基因组拷贝,并对两者进行了测序。该cDNA几乎是全长的,包含推定的翻译起始和终止密码子。概念翻译表明,grg-1 mRNA可指导合成一种分子量为7000的多肽。基因组克隆包含在一个1888 bp的PvuII片段中,涵盖了整个cDNA以及838 bp的5'侧翼序列和369 bp的3'侧翼序列。cDNA和基因组克隆的比较揭示了在潜在蛋白质编码序列中存在两个短内含子。发现grg-1的信使水平在葡萄糖剥夺开始后的几分钟内增加,并在去阻遏的前90分钟内上升50倍。