Salmikangas P, Keränen M R, Pajunen A
Biocenter, University of Oulu, Finland.
FEBS Lett. 1989 Nov 20;258(1):123-6. doi: 10.1016/0014-5793(89)81631-x.
The cDNA coding for rat S-adenosylmethionine decarboxylase (AdoMetDC, EC 4.1.1.50) has been cloned into a plasmid expression vector, pKK-223-3, and expressed in E. coli. The authenticity of the expressed protein has been demonstrated by reactivity with antibodies specific for rat AdoMetDC, by size analysis on SDS gels visualized with immunotransblots, and, finally, by catalytic activity. The expression of the enzyme results in a decrease in the activity of the bacterial enzyme suggesting the replacement of the bacterial enzyme by the rat AdoMetDC. Similarly, the addition of exogenous spermidine to the growth medium reduces bacterial enzyme activity affecting only marginally the expression of the recombinant protein.
编码大鼠S-腺苷甲硫氨酸脱羧酶（AdoMetDC，EC 4.1.1.50）的cDNA已被克隆到质粒表达载体pKK-223-3中，并在大肠杆菌中表达。通过与大鼠AdoMetDC特异性抗体的反应性、免疫印迹可视化的SDS凝胶上的大小分析以及最终的催化活性，证明了表达蛋白的真实性。该酶的表达导致细菌酶活性降低，表明大鼠AdoMetDC替代了细菌酶。同样，向生长培养基中添加外源亚精胺会降低细菌酶活性，对重组蛋白的表达仅产生轻微影响。
