克氏锥虫抗原多肽的体外合成

In vitro synthesis of Trypanosoma cruzi antigenic polypeptides.

作者信息

Stolf A M, Santos C L, Brentani R, Camargo M E

出版信息

Infect Immun. 1983 Mar;39(3):1175-9. doi: 10.1128/iai.39.3.1175-1179.1983.

DOI:10.1128/iai.39.3.1175-1179.1983

PMID:6404819

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC348080/

Abstract

A ribosomal fraction of Trypanosoma cruzi was isolated with detergent lysis and differential ultracentrifugation. This ribosomal fraction directed in vitro protein synthesis, in a heterologous incorporation system (rat liver pH 5 fraction), leading to values up to 10 times higher than endogenous control. The ideal concentrations of Mg2+, K+, and ribosomal proteins and the time of incubation of the incorporation mixtures were 6 mM, 21 mM, 60 microliters, and 45 min, respectively. The product thus obtained was analyzed by fluorography after polyacrylamide-sodium dodecyl sulfate gel electrophoresis and showed the presence of many protein bands, but few bands were present in the immunoprecipitate obtained with serum from Chagas' disease patients. This product was able to react with anti-T. cruzi antibodies when submitted to an immunoenzymatic assay.

摘要

通过去污剂裂解和差速超速离心法分离出克氏锥虫的核糖体部分。该核糖体部分在异源掺入系统（大鼠肝脏pH 5部分）中指导体外蛋白质合成，其合成值比内源性对照高10倍。掺入混合物中Mg2 +、K +、核糖体蛋白的理想浓度以及孵育时间分别为6 mM、21 mM、60微升和45分钟。聚丙烯酰胺 - 十二烷基硫酸钠凝胶电泳后，通过荧光自显影分析所得产物，结果显示存在许多蛋白条带，但恰加斯病患者血清免疫沉淀得到的条带较少。当进行免疫酶测定时，该产物能够与抗克氏锥虫抗体发生反应。