植物病毒载体介导的可溶性甲烷单加氧酶B亚单位在蚕豆植物中的表达

用基因操作技术将编码可溶性甲烷单加氧酶B亚单位（Soluble Methane Monoxygermse-B component：SMMO-B）的基因全长序列导入来自三叶草黄脉病毒（Clover yellow vein virus；CIYVV）侵染性全长cDNA克隆的侵染性植物病毒表达载体pCIYVV／CP／W基因组的NIb／CP基因之间，构建了重组病毒克隆pCV—mmoB。用上述重组病毒克隆接种该病毒的自然寄主植物蚕豆，表现了与野生型CIYVV相同的症状。RT—PCR检测结果表明，子代病毒基因组中的外源基因获得了转录。蛋白质杂交（Western blotting；WB）检测结果表明，sMMO—B亚单位蛋白在被重组病毒克隆侵染的寄主植物中获得了表达。

Expression of the sMMO-B in the Broad Bean Plant by the Plant Virus Vector

The sMMO-B was expressed in the broad bean plants by Potyvirus expression vector pClYVV/CP/W.The sequence of 420bp coding sMMO-B was inserted between nuclear inclusion b (NIb) and coat protein (CP) of infectious viral vector pClYVV/CP/W derived from clover yellow vein virus (ClYVV) of potyvirus.The recombinant plasmids were constructed and named pCV-MMO-B, and then they were mechanically inoculated in broad bean seedlings. The similar apparent symptom on the plant was observed post infection (pi) of the recombinant plasmids as well as the wild-type ClYVV.The genetic stability of the recombinant plasmids carrying foreign gene was examined by RT-PCR. The expressed sMMO-B protein were confirmed by western blot analysis using antibodies against the sMMO-B protein.The results showed that these proteins post expressing and automatically separated from the ClYVV pro-proteins were stable even in following subsequential passages of the progeny recombinants in broad bean plants.