Laperche Y, Bulle F, Aissani T, Chobert M N, Aggerbeck M, Hanoune J, Guellaën G
Proc Natl Acad Sci U S A. 1986 Feb;83(4):937-41. doi: 10.1073/pnas.83.4.937.
We have screened a cDNA library (20,000 clones) made from rat kidney poly(A)+ RNA, using an oligonucleotide probe that was a mixture of 14-base DNA oligomers containing all 32 possible sequences coding for residues 32-36 of the gamma-glutamyl transpeptidase (EC 2.3.2.2.) heavy chain. We isolated and sequenced two cDNAs corresponding to the mRNA coding for the entire length of the enzyme precursor. The nucleotide sequence that we obtained (2072 bases) reveals an open reading frame of 1707 nucleotides coding for the common precursor of both enzyme subunits. The amino acid sequence begins with the 21 residues located at the NH2-terminal hydrophobic region of the heavy subunit. We show that this sequence, which is not processed, is the only possible signal peptide in the sequence. Five potential N-glycosylation sites are present in the gamma-glutamyl transpeptidase sequence. Using one of the two cDNA clones as probe, a 2.2-kilobase sequence was detected by blot analysis in rat kidney and human fetal liver RNA.
我们使用一种寡核苷酸探针筛选了由大鼠肾脏多聚腺苷酸加尾RNA构建的cDNA文库（20,000个克隆），该探针是14碱基DNA寡聚物的混合物，包含编码γ-谷氨酰转肽酶（EC 2.3.2.2.）重链32 - 36位残基的所有32种可能序列。我们分离并测序了两个与编码该酶前体全长的mRNA相对应的cDNA。我们获得的核苷酸序列（2072个碱基）揭示了一个1707个核苷酸的开放阅读框，编码两种酶亚基的共同前体。氨基酸序列从位于重亚基NH2末端疏水区域的21个残基开始。我们表明，该未加工的序列是该序列中唯一可能的信号肽。γ-谷氨酰转肽酶序列中有五个潜在的N-糖基化位点。使用两个cDNA克隆之一作为探针，通过印迹分析在大鼠肾脏和人胎儿肝脏RNA中检测到一个2.2千碱基的序列。
