Neilson J T
J Parasitol. 1975 Oct;61(5):785-93.
A soluble somatic preparation (SSP) of adult Dipetalonema viteae was prepared. Aliquots of the aqueous insoluble debris (cuticles and membranes) left after the extraction of SSP were solubilized separately with Triton X-100 and lithium diiodosalicylate to yield a Triton solubilized preparation (TSP) and lithium diiodosalicylate solubilized preparation (LSP), respectively. SSP was sequentially chromatographed on a series of Sephadex columns, G50; then fraction 1 from G50 on G100 and finally fraction 1 from G100 on G200 yielding 5, 4, and 5 fractions on G50, G100, and G200, respectively. TSP and LSP were each sequentially chromatographed on Sephadex G50 and G200 to yield 5 and 4 fractions for TSP and 6 and 3 fractions for LSP, respectively. Each Sephadex fraction from each separation was further resolved by polyacrylamide gel electrophoresis (PAGE). Immunodiffusion studies with a rabbit antiserum to SSP gave 4 precipitin bands with SSP, 1 with TSP, and none with LSP. The single precipitin arc produced with TSP showed partial identity with an SSP arc. Disc immunoelectrophoresis allowed the identification of the proteins separated by PAGE, which precipitated with the rabbit anti-SSP serum.
制备了成年维氏双瓣线虫的可溶性虫体制剂(SSP)。在提取SSP后留下的水不溶性碎片(角质层和膜)的等分试样分别用 Triton X - 100 和二碘水杨酸锂溶解,分别得到 Triton 溶解制剂(TSP)和二碘水杨酸锂溶解制剂(LSP)。SSP 依次在一系列 Sephadex 柱(G50)上进行层析;然后将 G50 的第 1 级分在 G100 上进行层析,最后将 G100 的第 1 级分在 G200 上进行层析,在 G50、G100 和 G200 上分别得到 5、4 和 5 个级分。TSP 和 LSP 各自依次在 Sephadex G50 和 G200 上进行层析,TSP 分别得到 5 个和 4 个级分,LSP 分别得到 6 个和 3 个级分。每次分离得到的每个 Sephadex 级分通过聚丙烯酰胺凝胶电泳(PAGE)进一步分离。用兔抗 SSP 血清进行的免疫扩散研究表明,SSP 产生 4 条沉淀带,TSP 产生 1 条沉淀带,LSP 未产生沉淀带。TSP 产生的单一沉淀弧与 SSP 的一条弧显示部分同一性。圆盘免疫电泳可鉴定通过 PAGE 分离的、能与兔抗 SSP 血清沉淀的蛋白质。