Zhao D, Dai Z, Zhou K, Zhang L
College of Life Sciences, Nanjing Normal University, Nanjing 210097, China.
Wei Sheng Wu Xue Bao. 2001 Dec;41(6):680-5.
The synthesized CMIV-like Gene was linked with the signal peptide gene of nature silkworm antibacterial peptide and was inserted into baculovirus expression vector pFastBac 1, construcing a recombinant transposing vector. The vector was transformed into DH10Bac competent E. coli cells. The recombinant Bacmid was obtained. The recombinant Bacmid was transfected into sf21 cells to get the recombinant virus. Laphygma exigua larvae were infected with the recombinant virus to express the antibacterial peptide. The hemolymph were tested to have antibacterial activity. The active antibacterial peptide was purified by acid polyacrylamide electrophoresis. The specific expression of mRNA of CMIV-like Gene was tested using Northern blotting.
将合成的类CMIV基因与天然家蚕抗菌肽信号肽基因连接，插入杆状病毒表达载体pFastBac 1，构建重组转座载体。将该载体转化到DH10Bac感受态大肠杆菌细胞中，获得重组杆粒。将重组杆粒转染到sf21细胞中得到重组病毒。用重组病毒感染甜菜夜蛾幼虫以表达抗菌肽。检测血淋巴具有抗菌活性。通过酸性聚丙烯酰胺电泳纯化活性抗菌肽。使用Northern印迹法检测类CMIV基因mRNA的特异性表达。
Zotero 插件