Schudy A, Bangsow T, Flach R, Trost C, Schepelmann S, Weber P, Knüpfer E, Gassen H G
Institut für Biochemie, Technische Hochschule Darmstadt, Petersenstrasse 22, Darmstadt, D-64287, Germany.
Biochem Biophys Res Commun. 1996 Dec 24;229(3):693-700. doi: 10.1006/bbrc.1996.1867.
Three different mRNAs coding for the porcine gamma-glutamyl transpeptidase (GGT) in the kidney were identified by 5'-RACE-PCR. These differ in their 5'-noncoding region. Genomic Southern blot analysis has demonstrated the existence of a single GGT gene in the porcine genome. Thus, the existence of multiple mRNAs can only be explained by the use of different promoters or alternative splicing. Four GGT-specific genomic clones containing the complete 5'-end of the gene were isolated and characterized, revealing six exons common to all three mRNAs. Four of these exons were located in the coding region comprising the codons for amino acids 1 to 138. Two exons and an intervening sequence were identified upstream from these six common exons representing the unique 5'-ends of the three mRNAs. The coding exons show a significant sequence homology to mouse, rat, and human GGT cDNA, whereas exons 1 and 3 display no homology.
通过5'-RACE-PCR技术在猪肾中鉴定出三种编码猪γ-谷氨酰转肽酶(GGT)的不同mRNA。它们在5'-非编码区存在差异。基因组Southern印迹分析表明猪基因组中存在单一的GGT基因。因此,多个mRNA的存在只能通过使用不同的启动子或选择性剪接来解释。分离并鉴定了四个包含该基因完整5'-末端的GGT特异性基因组克隆,发现所有三种mRNA共有六个外显子。其中四个外显子位于编码区,包含氨基酸1至138的密码子。在这六个共有外显子上游鉴定出两个外显子和一个间隔序列,它们代表三种mRNA独特的5'-末端。编码外显子与小鼠、大鼠和人GGT cDNA具有显著的序列同源性,而外显子1和3则无同源性。