Moelans I I, Meis J F, Kocken C, Konings R N, Schoenmakers J G
Department of Molecular Biology, Faculty of Science, University of Nijmegen, The Netherlands.
Mol Biochem Parasitol. 1991 Apr;45(2):193-204. doi: 10.1016/0166-6851(91)90086-l.
A Plasmodium falciparum cDNA clone was isolated of which the insert is transcribed at high rates as a 1.4-kb mRNA in the sexual stages of the malaria parasite. The cDNA clone contains a copy of a non-interrupted gene which codes for a protein of 157 amino acids (Mr = 16607). This 16-kDa protein does not contain repetitive sequences and is characterised by a putative N-terminal signal sequence, a hydrophobic membrane anchor sequence and a highly hydrophilic C-terminal region suggesting that it is an integral membrane protein. Rabbit antisera raised against a synthetic peptide covering amino acids 31-47 of the 16-kDa protein and against recombinant fusion proteins recognised the 16-kDa antigen in protein extracts of gametocytes, macrogamete/zygotes and sporozoites by Western blot analysis. The rabbit antisera also reacted with gametes, gametocytes and sporozoites in a standard immunofluorescence assay. By immunoelectron microscopy using the protein A-gold method the 16-kDa protein could be clearly visualised on the surface of macrogametes and sporozoites, whereas the antigen was not detectable in the asexual erythrocytic stages of the parasite. The 16-kDa antigen of P. falciparum therefore might have the potential to elicit a dual protective immune response against the sporozoite and sexual stage parasites.
分离出一个恶性疟原虫cDNA克隆,其插入片段在疟原虫有性阶段以1.4kb mRNA的形式进行高转录。该cDNA克隆包含一个不间断基因的拷贝,该基因编码一种由157个氨基酸组成的蛋白质(Mr = 16607)。这种16kDa的蛋白质不包含重复序列,其特征在于一个推定的N端信号序列、一个疏水膜锚定序列和一个高度亲水的C端区域,表明它是一种整合膜蛋白。用针对覆盖16kDa蛋白质氨基酸31 - 47的合成肽以及重组融合蛋白产生的兔抗血清,通过蛋白质印迹分析在配子体、大配子/合子和子孢子的蛋白质提取物中识别出16kDa抗原。在标准免疫荧光试验中,兔抗血清也与配子、配子体和子孢子发生反应。通过使用蛋白A - 金法的免疫电子显微镜观察,可在大配子和子孢子表面清晰地看到16kDa蛋白质,而在该寄生虫的无性红细胞阶段未检测到该抗原。因此,恶性疟原虫的16kDa抗原可能有潜力引发针对子孢子和有性阶段寄生虫的双重保护性免疫反应。