Mowatt M R, Weinbach E C, Howard T C, Nash T E
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Exp Parasitol. 1994 Feb;78(1):85-92. doi: 10.1006/expr.1994.1008.
The Giardia lamblia triosephosphate isomerase (TIM) gene was cloned using a probe generated by a polymerase chain reaction that employed primers complementary to highly conserved regions of TIM. The nucleotide sequence predicts a protein that is 38 and 47% identical to TIM from prokaryotic and eukaryotic sources, respectively. Like all other Giardia protein-coding genes studied thus far, the TIM gene lacks introns and is transcribed to yield a polyadenylated mRNA with an extremely short 5' untranslated region. The Giardia TIM gene complemented an Escherichia coli triosephosphate isomerase deletion mutant. The simplicity and success of complementation suggests its general utility in cloning Giardia genes of known function.
利用聚合酶链反应生成的探针克隆了蓝氏贾第鞭毛虫磷酸丙糖异构酶（TIM）基因，该聚合酶链反应使用了与TIM高度保守区域互补的引物。核苷酸序列预测的一种蛋白质，与来自原核生物和真核生物来源的TIM分别有38%和47%的同一性。与迄今为止研究的所有其他贾第鞭毛虫蛋白质编码基因一样，TIM基因缺乏内含子，转录后产生一个5'非翻译区极短的多聚腺苷酸化mRNA。蓝氏贾第鞭毛虫TIM基因补充了大肠杆菌磷酸丙糖异构酶缺失突变体。互补的简易性和成功表明其在克隆已知功能的贾第鞭毛虫基因方面具有普遍用途。
