AMINO ACID AT THE SUPPRESSION SITE IN RABBIT BETA-GLOBIN READTHROUGH PROTEIN

兔β-珠蛋白通读蛋白是唯一已知的天然 存在于高等真核生物中的通读蛋白， 涉及病毒系统。 由于抑制性tRNA已被用于 基因治疗实验，并已涉及抑制 病毒表达，重要的是要表征的无意义 抑制tRNA参与这种独特蛋白质的表达。 因此，设计了实验来分离这种蛋白质， 兔网织红细胞，并确定氨基酸在 通读网站。 针对这种蛋白质的特异性抗体， 通过合成22个氨基酸的肽来制备， 对于β-珠蛋白通读蛋白的通读部分， 将所述肽偶联至KLH蛋白，并将所述肽注射至所述KLH蛋白， 结合蛋白质注入绵羊体内 产生了特异性抗体 将其纯化并用于分离通读蛋白。 蛋白质通过电泳纯化至均一， 聚丙烯酰胺凝胶和纯化的产品目前正在 通过通读网站进行排序。

Rabbit beta-globin readthrough protein is the only known naturally occurring readthrough protein in higher eukaryotes which does not involve a viral system. Since suppressor tRNAs have been used in gene therapy experiments and have been implicated in inhibiting viral expression, it is important to characterize the nonsense suppressor tRNA involved in the expression of this unique protein. Experiments were devised, therefore, to isolate this protein from rabbit reticulocytes and to identify the amino acid at the readthrough site. Specific antibodies against this protein were prepared by synthesizing a 22 amino acid peptide which corresponds to the readthrough portion of the beta-globin readthrough protein, coupling the peptide to the KLH protein and injecting the conjugated protein into a sheep. Specific antibodies were produced which were purified and used to isolate the readthrough protein. The protein was purified to homogeneity by electrophoresis on a polyacrylamide gel and the purified product is presently being sequenced through the readthrough site.