Kutchan T M, Hampp N, Lottspeich F, Beyreuther K, Zenk M H
Lehrstuhl für Pharmazeutische Biologie, Universität München, FRG.
FEBS Lett. 1988 Sep 12;237(1-2):40-4. doi: 10.1016/0014-5793(88)80167-4.
The cDNA clone for strictosidine synthase, the enzyme which catalyzes the stereospecific condensation of tryptamine with secologanin to form the key intermediate in indole alkaloid biosynthesis, strictosidine, has been identified with a synthetic oligodeoxynucleotide hybridization probe in a lambda gt11 cDNA library of cultured cells of Rauvolfia serpentina. The DNA has been sequenced, revealing an open reading frame of 1032 base pairs encoding 344 amino acids. The sequence of 60 nucleotides in the 5'-flanking region has been determined by primer extension analysis. The encoded protein has been expressed in E. coli DH5 as detected by immunoblotting of protein extracts with antibodies raised against the native enzyme.
已使用合成的寡聚脱氧核苷酸杂交探针，在蛇根木培养细胞的λgt11 cDNA文库中鉴定出了用于催化色胺与番木鳖苷元立体特异性缩合形成吲哚生物碱生物合成关键中间体——士的宁苷的裂环马钱子苷合酶的cDNA克隆。该DNA已测序，揭示了一个1032个碱基对的开放阅读框，编码344个氨基酸。通过引物延伸分析确定了5'侧翼区域60个核苷酸的序列。如用针对天然酶产生的抗体对蛋白质提取物进行免疫印迹检测所示，编码的蛋白质已在大肠杆菌DH5中表达。
