Integration of Non-natural Amino Acids Into Protein

1)为了获得较高的氨基酰化pCpCpA的产率，对5‘端和核苷酸间磷酸及碱基胺的几种保护基团进行了实验，设计合成了生物素赖氨基PCPA，使蛋白质整合后的氨基酸检测变得更容易。2)利用显微外科技术和酵母tRNA^&lt、Phe&gt；、tRNA^&lt、Tyr&GT等，建立了一种高效的制备具有任意核苷酸序列反密码子的tRNA的方法。在此基础上，建立了一种非放射性的低分子RNA核苷酸序列测定方法。3)在大肠杆菌无细胞翻译系统中，硫代硫代信使核糖核酸(硫代信使核糖核酸)比天然信使核糖核酸具有更强的抗核酸酶活性和翻译效率。在极端嗜热菌HB8的无细胞体系中，研究了不寻常的多胺对苯丙氨酰-tRNA形成的影响。此外，还研究了酵母、圆柱念珠菌和牛线粒体细胞-树翻译系统的基本特征。

1) Several protecting groups of 5'-terminal and internucleotide phosphates and base amines were tested to obtain better yield of aminoacyl pCpCpA.Biotinyl-lysyl pCpA was designed and synthesized to make it easier to detect the amino acid after integration into proteins.2) An efficient method for preparing tRNA which has an anticodon of arbitrary nucleotide sequence and lacks 3'-terminal pCpCpA,was developed by the use of mictosurgery technique and yeast tRNA^<Phe>, tRNA^<Tyr> and so on. Furthermore, a non-radioactive method for determining nucleotide sequences of low-molecular-weight RNA was developed.3) Advantage in nuclease resistance and translational efficiency of phosphorothioate-containing mRNA (thio-mRNA) over natural mRNA was demonstrated in E.coli cell-free translation system. Effects of unusual polyamines on phenylalanyl-tRNA formation was investigated in cell-free systems of an extreme thermophile, Thermus themophilus HB8. Fundamental characteristics of cell-tree translation systems from yeast, Candida cylindracea and bovine mitochondriawere also examined.