Project Directorate on Animal Disease Monitoring and Surveillance (PD_ADMAS), Hebbal, Bengaluru, Karnataka 560024, India. pinakiprasad
Vet Parasitol. 2012 Jun 8;187(1-2):1-8. doi: 10.1016/j.vetpar.2012.01.012. Epub 2012 Jan 10.
The variant surface glycoprotein (VSG) of trypanosome is an important part of its body surface coat, which is expressed in early, middle and late stages of infection contributing a major diagnostic value. In the present study, the 5' end of the partial VSG gene sequences (681 bp) encoding N-terminal protein of RoTat 1.2 VSG (227 amino acid) was amplified, cloned into pET32a vector, and expressed in prokaryotic system. The fused His-tagged expressed VSG protein (43 kDa) of the Trypanosoma evansi was characterized in SDS-PAGE and immunoblotting using hyperimmune/immune sera raised against buffalo, dog, lion and leopard isolates of T. evansi. The expressed protein remained immunoreactive with all the sera combinations. The animals immunized with whole cell lysate or recombinant protein showed similar antibody reactions in ELISA and CATT (Card Agglutination Test for Trypanosomiasis). This study suggests the expressed recombinant truncated VSG is having its importance for its possible use in sero-diagnosis of surra.
锥虫的变异表面糖蛋白（VSG）是其体表面被膜的重要组成部分，在感染的早期、中期和晚期表达，具有重要的诊断价值。本研究扩增了编码 RoTat 1.2 VSG（227 个氨基酸）N 端蛋白的部分 VSG 基因序列（681bp），克隆到 pET32a 载体中，并在原核系统中表达。用针对水牛、狗、狮子和豹分离株的抗锥虫伊氏锥虫的高免/免疫血清在 SDS-PAGE 和免疫印迹中对表达的融合 His 标记的 VSG 蛋白（43kDa）进行了鉴定。表达的蛋白与所有血清组合保持免疫反应性。用全细胞裂解物或重组蛋白免疫的动物在 ELISA 和 CATT（锥虫病卡片凝集试验）中表现出相似的抗体反应。这项研究表明，表达的截短的重组 VSG 具有重要意义，可能用于苏拉病的血清学诊断。
