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编码在虫卵胚胎发生过程中表达的绵羊带绦虫疫苗抗原的基因家族的序列分析。

Sequence analysis of a gene family encoding Taenia ovis vaccine antigens expressed during embryogenesis of eggs.

作者信息

Waterkeyn J, Gauci C, Cowman A, Lightowlers M

机构信息

University of Melbourne, Werribee, Victoria, Australia.

出版信息

Mol Biochem Parasitol. 1997 May;86(1):75-84. doi: 10.1016/s0166-6851(97)90007-4.

Abstract

The 45W vaccine antigen of the cestode parasite Taenia ovis is a member of a small gene family estimated to comprise six members. All six genes were cloned and characterised. Nucleotide sequence analysis of the different family members identified a set of closely related genes exhibiting between 75.2 and 98.6% DNA sequence homology. Intron/exon structure for the various genes constituting the family was found to be conserved. Open reading frame analysis and pairwise alignments of predicted polypeptides from the various members of the family revealed that the encoded proteins share between 51.8 and 96.5% amino acid identity with the host-protective 45W antigen. Polymerase chain reaction (PCR) primers were designed which allowed amplification of individual family members in a transcript-specific manner. These were used to investigate gene expression in different developmental stages of the parasite by reverse-transcription polymerase chain reaction (RT-PCR). All members of the gene family were expressed during the T. ovis life-cycle. 45W gene family transcription correlates with embryogenesis of eggs and expression is increased in activated eggs (oncospheres). One family member, ToW3S, was expressed in the early larvae (cysticercus) stages. Expression of genes closely related to the 45W vaccine antigen has important implications for immune evasion under selection from the application of a 45W-based vaccine.

摘要

绦虫寄生虫绵羊带绦虫的45W疫苗抗原是一个小基因家族的成员,估计该家族由六个成员组成。所有六个基因都被克隆并进行了表征。对不同家族成员的核苷酸序列分析确定了一组密切相关的基因,它们的DNA序列同源性在75.2%至98.6%之间。发现构成该家族的各种基因的内含子/外显子结构是保守的。对该家族各个成员预测的多肽进行开放阅读框分析和两两比对,结果显示,编码的蛋白质与具有宿主保护作用的45W抗原的氨基酸同一性在51.8%至96.5%之间。设计了聚合酶链反应(PCR)引物,可用于以转录本特异性方式扩增各个家族成员。通过逆转录聚合酶链反应(RT-PCR),利用这些引物研究寄生虫不同发育阶段的基因表达。该基因家族的所有成员在绵羊带绦虫的生命周期中均有表达。45W基因家族的转录与虫卵的胚胎发生相关,并且在活化的虫卵(六钩蚴)中表达增加。一个家族成员ToW3S在幼虫早期(囊尾蚴)阶段表达。与45W疫苗抗原密切相关的基因的表达对于在基于45W的疫苗应用选择下的免疫逃避具有重要意义。

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