Molecular analysis of the gene encoding the major surface antigen of Toxoplasma gondii.
Molecular analysis of the gene encoding the major surface antigen of Toxoplasma gondii.
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DOI：
10.4049/jimmunol.141.10.3584
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发表时间：
1988-11
影响因子：
4.4
通讯作者：
J. L. Burg;D. Perelman;L. Kasper;P. Ware;J. Boothroyd
中科院分区：
文献类型：
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作者：
J. L. Burg;D. Perelman;L. Kasper;P. Ware;J. Boothroyd
通过对弓形虫主要表面抗原P30基因的克隆和分析，推导出了P30基因的全序列。利用P30特异性多克隆血清，我们从T.弓形虫RH株。该克隆产生与几种抗P30 mAb反应的β-半乳糖苷酶融合蛋白。此外，针对融合蛋白的多克隆抗血清与纯化的P30蛋白反应，并且仅与T.刚地该cDNA克隆用于分离近全长cDNA分子和含有P30基因的粘粒克隆。cDNA的序列分析揭示了一个单一的开放阅读框，其编码能力为34.7 kDa的初级翻译产物（与SDS-丙烯酰胺凝胶上P30的表观Mr一致），包括一个假定的疏水信号序列。P30初级翻译产物还具有羧基末端疏水尾，其预测翻译后裂解和糖脂锚修饰。我们已经鉴定了P30 mRNA转录物的表观5'和3'末端，其非常丰富，长度为1500个核苷酸，并且是多腺苷酸化的。P30基因是单拷贝的，不含内含子。
The complete sequence of P30, the major surface Ag of the protozoan parasite, Toxoplasma gondii, has been deduced through the cloning and analysis of its gene. Using polyclonal serum specific for P30, we have isolated a P30 cDNA clone from a lambda gt11 cDNA expression library derived from tachyzoites of T. gondii (RH strain). This clone produces a beta-galactosidase fusion protein which reacts with several anti-P30 mAb. In addition, polyclonal anti-serum raised to the fusion protein reacts with purified P30 protein and exclusively with P30 in a whole cell lysate of T. gondii. This cDNA clone was used to isolate near full-length cDNA molecules and a cosmid clone containing the P30 gene. Sequence analysis of the cDNA reveals a single open reading frame with coding capacity for 34.7 kDa of primary translation product (consistent with the apparent Mr of P30 on SDS-acrylamide gels) including a presumptive hydrophobic signal sequence. The P30 primary translation product also has a carboxy-terminal hydrophobic tail which is predictive of a posttranslational cleavage and modification with a glycolipid anchor. We have identified the apparent 5' and 3' ends of the P30 mRNA transcript which is extremely abundant, 1500 nucleotides in length, and polyadenylated. The P30 gene is single copy and contains no introns.