人源抗菌肽LL-37在烟草中的表达

为探讨用植物生物反应器生产LL-37的可行性,利用DNA重组技术将LL-37与植物病程相关蛋白信号肽基因融合,经测序证实序列和阅读框正确后,连接到质粒pBin438上,构建含LL-37 cDNA植物分泌型表达载体pBin/LL-37.通过根癌土壤杆菌(Agrobacterium tumefacience)介叶盘法转化烟草(Nicotiana tobacum.NC-89).用PCR和Western blot对转化植株进行分子鉴定.80%转基因植株表达重组LL-37多肽.T0代转基因烟草的蛋白粗提物有抑制Staphylococcus aureus和Escherichia coli生长的活性.LL-37基因在转基因烟草中成功表达,提示用植物表达系统生产活性LL-37蛋白是可行的.

Expression of human antimicrobial peptide LL-37 in Nicotiana tabacum

A cDNA fragment encoding human cathelicidin LL-37 linked to a signal peptide sequence of tobacco pathogenesis-related protein 1b gene was introduced into a plant expression vector, pBin438 and stably integrated into Nicotiana tabacum cv. NC-89 genome using Agrobacterium tumefaciens-mediated transformation. The presence of the fusion cDNA in the transgenic plant was confirmed by PCR and its expression was confirmed by Western blot analysis. Protein extracts from transgenic tobacco plant leaves exhibited antibacterial activity. This is the first report of synthesis of biologically active LL-37 in plants.