用差异显示技术寻找家蚕诱导前后特异表达的基因

Searching for New Genes Involved in Immune Reactions of Bombyx mori Through Differential Display

分别抽取诱导和未诱导家蚕（Ｂｏｍｂｙｘｍｏｒｉ）蛹脂肪体总ＲＮＡ进行ＤＤＲＴＰＣＲ反应。产物于非变性聚丙烯酰胺凝胶上电泳展开，得到的差异条带扩增后大多可以在２％琼脂糖凝胶上得到一条以上的带。Ｎｏｒｔｈｅｒｎｂｌｏｔ证明为阳性的一条带克隆后得到６个阳性转化子，ＳＳＣＰ证明它们含有相同的插入片段。对其中之一进行测序，结果显示片段长２４１ｂｐ，没有发现同源性较高的基因。对此基因的进一步研究有望揭示它在昆虫免疫反应中可能扮演的角色。

Total RNAs of induced and non induced Bombyx mori pupa fat bodies are purified respectively and then subjected to DDRT PCR. After displayed on native polyacrylamide gels, eight differentially expressed bands are retrieved and reamplified, most of which include more than one band when visualized in 2% agarose gels. One fragment proved to be positive in Northern blot is cloned and six positive recombinants are proved to have the same insert by SSCP. Sequencing result shows it is 241bp long and no homologies are found. Further studies may reveal it possible role in insect immune reactions.