Liu J, Koltai H, Chejanovsky N, Spiegel Y
Department of Nematology, Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel.
J Parasitol. 2001 Aug;87(4):801-7. doi: 10.1645/0022-3395(2001)087[0801:IOANCG]2.0.CO;2.
Mj-col-5, isolated from the plant parasitic nematode Meloidogyne javanica, has a longer carboxy-terminus than other members of the Caenorhabditis elegans COL-6 subfamily of cuticle collagen, including an extra tyrosine residue, and may form altered nonreducible cross-linkages. By semiquantitative determination at different life stages, Mj-col-5 transcript was shown to be more abundant in eggs than in juveniles/young females and adult females. To characterize further this gene's contribution to the changing cuticle of the nematode, we expressed a fusion protein containing a nonconserved 58-amino-acid sequence from the putative Mj-col-5 gene product and raised rabbit antiserum against the fusion protein. The antiserum detected a strongly reacting band (36 kDa, designated MJE36) on western blots of M. javanica eggs extracted with beta-mercaptoethanol. MJE36 was sensitive to collagenase and was not detected on western blots of extracts from M. javanica second-stage juveniles or adult females. A band of the same molecular size was detected in Meloidogyne incognita egg extracts but not in those of Heterodera avenae. Immunoblot indicated that MJE36 is not present in egg shells of M. javanica.
从植物寄生线虫爪哇根结线虫（Meloidogyne javanica）中分离出的Mj-col-5，其羧基末端比秀丽隐杆线虫（Caenorhabditis elegans）表皮胶原蛋白COL-6亚家族的其他成员更长，包括一个额外的酪氨酸残基，并且可能形成改变的不可还原交联。通过在不同生命阶段的半定量测定，发现Mj-col-5转录本在卵中的丰度高于幼虫/年轻雌虫和成年雌虫。为了进一步表征该基因对线虫不断变化的表皮的作用，我们表达了一种融合蛋白，该融合蛋白包含来自推测的Mj-col-5基因产物的一个非保守的58个氨基酸序列，并制备了针对该融合蛋白的兔抗血清。该抗血清在用β-巯基乙醇提取的爪哇根结线虫卵的蛋白质免疫印迹上检测到一条强反应带（36 kDa，命名为MJE36）。MJE36对胶原酶敏感，在用β-巯基乙醇提取的爪哇根结线虫二期幼虫或成年雌虫的提取物的蛋白质免疫印迹上未检测到。在南方根结线虫（Meloidogyne incognita）卵提取物中检测到相同分子大小的条带，但在燕麦孢囊线虫（Heterodera avenae）的提取物中未检测到。免疫印迹表明MJE36不存在于爪哇根结线虫的卵壳中。
