| 中华蜜蜂前溶血肽原基因在Tn-5B1-4细胞中的表达 |
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| 引用本文： | 施婉君,张传溪,程家安.中华蜜蜂前溶血肽原基因在Tn-5B1-4细胞中的表达[J].上海交通大学学报(农业科学版),2005,23(2):117-121. |
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| 作者姓名： | 施婉君 张传溪 程家安 |
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| 作者单位： | 1. 浙江大学,应用昆虫学研究所,杭州,310029;上海交通大学,资源环境科学系,上海,201101
2. 浙江大学,应用昆虫学研究所,杭州,310029 |
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| 基金项目： | 上海交通大学院青年基金(AE150054)，国家自然基金(No.30271008) |
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| 摘 要： | 构建中华蜜蜂前溶血肽原基因的重组供体质粒pBacHT—GFPTAccPPM，将其转化进入穿梭栽体Bacmid的受体茵DH10Bac中，得重组穿梭栽体Bacmid-GFPTAccPPM，再用Lipofectin介导其DNA转染粉纹夜蛾细胞系Tn-581-4，最后观察细胞表达时相动态。经ELISA法和荧光显微镜观察证实，胞内表达产物既能发射易于检测的绿色荧光，又具有良好的抗原性，证明重组病毒Bacmid-GFPTAccPPM已在昆虫细胞中得到了表达。
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| 关 键 词： | 中华蜜蜂 基因 溶血 粉纹夜蛾细胞系 ELISA法 肽 DNA转染 显微镜观察 细胞表达 绿色荧光 表达产物 昆虫细胞 重组病毒 抗原性 栽体 穿梭 供体 介导 |
| 文章编号： | 1671-9964(2005)02-0117-05 |
| 修稿时间： | 2004年12月15 |
Expression of Prepromelittin Gene of Apis cerana cerana in Tn-5B1-4 cell |
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| SHI Wan-jun,ZHANG Chuan-xi,CHENG Jia-an.Expression of Prepromelittin Gene of Apis cerana cerana in Tn-5B1-4 cell[J].Journal of Shanghai Jiaotong University (Agricultural Science),2005,23(2):117-121. |
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| Authors: | SHI Wan-jun ZHANG Chuan-xi CHENG Jia-an |
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| Institution: | SHI Wan-jun1,2,ZHANG Chuan-xi1,CHENG Jia-an 1 |
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| Abstract: | The cDNA encoding prepromelittin in Apis cerana cerana was obtain ed by restriction enzyme digestion from the recombinant plasmid and inserted into the multiple cloning site of the pBacHTeGFPT to construct recombinant donor plas mid, pBacHT- GFPTAccPPM, which was used to transposed to the target Bacmid in E . coli (DH10) by Tn7 transposition function. Then the recombinant Bacmids were t ransfected into Tn- 5B1- 4 cells of the cabbage looper, Trichoplusia ni, media ted by Lipofectin. The expressed protein detected by ELISA and the luminescence observed under the fluorescent microscope indicate that the recombinant bacmid h as been expressed in Tn-5B1-4 cell. |
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| Keywords: | Apis cerana cerana prepromelittin expression |
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