A transgenic silkworm expressing the immune-inducible cecropin B-GFP reporter gene
A transgenic silkworm expressing the immune-inducible cecropin B-GFP reporter gene
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DOI：
10.1016/j.ibmb.2006.03.002
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发表时间：
2006-05-01
影响因子：
3.8
通讯作者：
Taniai, Kiyoko
中科院分区：
文献类型：
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作者：
Imamura, Morikazu;Nakahara, Yuichi;Taniai, Kiyoko
为了分析天蚕素 B 启动子 (P-CecB) 体内活性，我们使用 PiggyBac 转座元件构建了在 P-CecB 控制下表达 EGFP 的转基因蚕。 G1 和 G2 代的基因组 Southern 印迹分析表明 EGFP 在基因组中稳定插入。将大肠杆菌细胞注射到幼虫体内，强烈诱导脂肪体和所有五种血细胞类型中的 EGFP 表达。 Northern印迹分析表明，细菌注射后脂肪体中EGFP的表达动力学与内源性CecB的表达动力学相关。对血细胞的流式细胞术分析表明，细菌（而非酵母）增加了 EGFP 表达。我们的结果表明，转基因蚕中 EGFP 表达的特征与内源性 CecB 的特征相同，并且可以通过使用转基因蚕的 EGFP 表达来监测 P-CecB 的激活。 (c) 2006 Elsevier Ltd. 保留所有权利。
To analyze cecropin B promoter (P-CecB) activity in vivo, we constructed transgenic silkworms that expressed EGFP under the control of P-CecB using the piggyBac transposable element. Genomic Southern blot analysis of the G1 and G2 generations indicated the stable insertion of EGFP in the genome. Injection of Escherichia coli cells into the larvae strongly induced EGFP expression in the fat bodies and all five hemocyte cell types. Northern blot analysis indicated that the expression kinetics of EGFP in the fat bodies following bacterial injection were correlated with that of endogenous CecB. Flow cytometric analysis of the hemocytes revealed that EGFP expression was increased by bacteria, but not by yeast. Our results indicate that the features of EGFP expression in the transgenic silkworm are equivalent to those of endogenous CecB and that P-CecB activation can be monitored by EGFP expression using transgenic silkworms. (c) 2006 Elsevier Ltd. All rights reserved.