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克氏锥虫的基因组包含一个与主要热休克蛋白同源的组成型表达、串联排列的多拷贝基因。

The genome of Trypanosoma cruzi contains a constitutively expressed, tandemly arranged multicopy gene homologous to a major heat shock protein.

作者信息

Dragon E A, Sias S R, Kato E A, Gabe J D

出版信息

Mol Cell Biol. 1987 Mar;7(3):1271-5. doi: 10.1128/mcb.7.3.1271-1275.1987.

Abstract

cDNA libraries have been constructed in the plasmid vector pUC18 with mRNA isolated from both epimastigotes and trypomastigotes of the Peru strain of Trypanosoma cruzi. Pools of randomly selected clones were analyzed by hybridization-selection-translation. Translation products were immunoprecipitated either with normal human sera or with sera from patients with Chagas' disease (chagasic sera), and the immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With this approach, a cDNA clone (pEC5) was identified which encodes a portion of an 85,000-Mr polypeptide. A genomic clone was subsequently isolated (FG1) by using oligonucleotide probes derived from the DNA sequence of this cDNA clone. A portion of this clone was isolated and sequenced, and the coding region for the protein was identified. Computer analysis of the predicted protein sequence indicates that this protein is closely related to the 83,000-Mr heat shock protein (hsp83) of Drosophila melanogaster, the hsp90 of Saccharomyces cerevisiae, and the hsp90 of chicken. This gene is tandemly organized in the T. cruzi genome as a cluster of 6 to 10 copies.

摘要

利用从克氏锥虫秘鲁株的上鞭毛体和锥鞭毛体中分离的mRNA,构建了pUC18质粒载体的cDNA文库。通过杂交选择翻译分析随机挑选的克隆池。用正常人血清或恰加斯病患者血清(恰加斯血清)对翻译产物进行免疫沉淀,并用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析免疫沉淀物。通过这种方法,鉴定出一个编码85000道尔顿多肽一部分的cDNA克隆(pEC5)。随后,利用从该cDNA克隆的DNA序列衍生的寡核苷酸探针分离出一个基因组克隆(FG1)。分离并测序该克隆的一部分,确定了该蛋白质的编码区。对预测的蛋白质序列进行计算机分析表明,该蛋白质与黑腹果蝇的83000道尔顿热休克蛋白(hsp83)、酿酒酵母的hsp90和鸡的hsp90密切相关。该基因在克氏锥虫基因组中串联排列,为6至10个拷贝的簇。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e58e/365202/af2af308695d/molcellb00075-0318-a.jpg

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