约氏疟原虫富含组氨酸蛋白的生物合成及其基因在大肠杆菌中的克隆。
The biosynthesis of the histidine-rich protein of Plasmodium lophurae and the cloning of its gene in Escherichia coli.
作者信息
Kilejian A, Chen S, Sloma A
出版信息
Mol Biochem Parasitol. 1985 Jan;14(1):1-10. doi: 10.1016/0166-6851(85)90100-8.
Uninucleate trophozoites and schizonts of Plasmodium lophurae were labeled metabolically with [3H]proline. Analysis of labeled parasites indicated that the histidine-rich protein (HRP) was the major polypeptide synthesized by both developmental stages; in trophozoites it represented a larger proportion of total labeled polypeptides. Polyadenylated RNA was prepared from trophozoites and translated in a rabbit reticulocyte lysate system in the presence of [3H]histidine. As compared to the approx. 50 kDa mature protein, in the cell-free system HRP was synthesized as an approx. 58 kDa precursor polypeptide. Size-selected, polyadenylated RNA was used to construct a complementary double-stranded DNA library in pBR322 and plasmids containing HRP sequences were identified by hybridization with a synthetic oligonucleotide probe.
用[3H]脯氨酸对鸡疟原虫的单核滋养体和裂殖体进行代谢标记。对标记寄生虫的分析表明，富含组氨酸的蛋白（HRP）是这两个发育阶段合成的主要多肽；在滋养体中，它在总标记多肽中所占比例更大。从滋养体中制备多聚腺苷酸化RNA，并在[3H]组氨酸存在的情况下在兔网织红细胞裂解物系统中进行翻译。与约50 kDa的成熟蛋白相比，在无细胞系统中，HRP是以约58 kDa的前体多肽形式合成的。经大小选择的多聚腺苷酸化RNA用于构建pBR322中的互补双链DNA文库，并通过与合成寡核苷酸探针杂交来鉴定含有HRP序列的质粒。
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