Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8144, USA.
FEBS Lett. 2012 Oct 19;586(20):3716-22. doi: 10.1016/j.febslet.2012.08.031. Epub 2012 Sep 13.
Genetically encoded phosphoserine incorporation programmed by the UAG codon was achieved by addition of engineered elongation factor and an archaeal aminoacyl-tRNA synthetase to the normal Escherichia coli translation machinery (Park et al., 2011) Science 333, 1151). However, protein yield suffers from expression of the orthogonal phosphoserine translation system and competition with release factor 1 (RF-1). In a strain lacking RF-1, phosphoserine phosphatase, and where seven UAG codons residing in essential genes were converted to UAA, phosphoserine incorporation into GFP and WNK4 was significantly elevated, but with an accompanying loss in cellular fitness and viability.
通过向正常的大肠杆菌翻译机制中添加工程化延伸因子和古菌氨酰-tRNA 合成酶，实现了由 UAG 密码子编码的磷酸丝氨酸掺入（Park 等人，2011 年）。然而，由于正交磷酸丝氨酸翻译系统的表达和与释放因子 1（RF-1）的竞争，蛋白质产量受到影响。在缺乏 RF-1、磷酸丝氨酸磷酸酶的菌株中，将七个位于必需基因中的 UAG 密码子转换为 UAA，GFP 和 WNK4 中的磷酸丝氨酸掺入显著增加，但伴随细胞适应性和活力的丧失。
