Piper P W, Emson C, Jones C E, Cowan D A, Fleming T M, Littlechild J A
Department of Biochemistry and Molecular Biology, University College London, London WC1E 6BT, UK.
Curr Genet. 1996 May;29(6):594-6.
The gene encoding phosphoglycerate kinase (PGK) from the Archaeon Sulfolobus solfataricus, an organism growing optimally at 87 degrees C, was inserted into a yeast expression vector under the control of the galactose-inducible GAL1 yeast promoter. This vector was then transformed into a pgk::TRP1 yeast mutant, a strain inhibited for growth on galactose or glucose due to its lack of PGK enzyme. Slow-growing transformants were obtained on galactose plates at 37 degrees C, but not 28 degrees C. These transformants contained low levels of transcripts of the heterologous gene and low amounts of thermostable PGK activity. Weak expression of the hyperthermophile gene in yeast, a mesophile, therefore enabled complementation of the yeast pgk defect at 37 degrees C but not at 28 degrees C.
编码磷酸甘油酸激酶（PGK）的基因来自嗜热栖热菌，这种生物在87摄氏度时生长最佳，该基因被插入到在半乳糖诱导型GAL1酵母启动子控制下的酵母表达载体中。然后将该载体转化到pgk::TRP1酵母突变体中，该菌株由于缺乏PGK酶而在半乳糖或葡萄糖上生长受到抑制。在37摄氏度的半乳糖平板上获得了生长缓慢的转化体，但在28摄氏度时未获得。这些转化体含有低水平的异源基因转录本和少量的热稳定PGK活性。因此，嗜热基因在中温菌酵母中的弱表达能够在37摄氏度而非28摄氏度时互补酵母pgk缺陷。
