Nagasu T, Hall B D
Gene. 1985;37(1-3):247-53. doi: 10.1016/0378-1119(85)90279-3.
The isolation of the Saccharomyces cerevisiae gene for NADP-dependent glutamate dehydrogenase (NADP-GDH) by cross hybridization to the Neurospora crassa am gene, known to encode for NADP-GDH is described. Two DNA fragments selected from a yeast genomic library in phage lambda gt11 were shown by restriction analysis to share 2.5 kb of common sequence. A yeast shuttle vector (CV13) carrying either to the cloned fragments complements the gdh- strain of S. cerevisiae and directs substantial overproduction of NADP-GDH. One of the cloned fragments was sequenced, and the deduced amino acid (aa) sequence of the yeast NADP-GDH is 64% homologous to N. crassa, 51% to Escherichia coli and 24% to bovine NADP-GDHs.
本文描述了通过与已知编码NADP依赖性谷氨酸脱氢酶(NADP-GDH)的粗糙脉孢菌am基因进行交叉杂交,分离酿酒酵母NADP-GDH基因的过程。从噬菌体λgt11中的酵母基因组文库中筛选出的两个DNA片段,经限制性分析表明共有2.5 kb的共同序列。携带任一克隆片段的酵母穿梭载体(CV13)可互补酿酒酵母的gdh-菌株,并大量过量表达NADP-GDH。对其中一个克隆片段进行了测序,酵母NADP-GDH推导的氨基酸(aa)序列与粗糙脉孢菌的同源性为64%,与大肠杆菌的为51%,与牛NADP-GDH的为24%。