Transformation of Internal Extracellular Bacteria Isolated from Rhagoletis completa Cresson Gut with Enhanced Green Fluorescent Protein
Transformation of Internal Extracellular Bacteria Isolated from Rhagoletis completa Cresson Gut with Enhanced Green Fluorescent Protein
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DOI：
10.1007/s002840010072
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发表时间：
2000-04
影响因子：
2.6
通讯作者：
J. Peloquin;L. Kuzina;C. Lauzon;T. Miller
中科院分区：
文献类型：
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作者：
J. Peloquin;L. Kuzina;C. Lauzon;T. Miller
我们发现 Zeocin™ 是一种有效的抗生素，可对抗从核桃壳蝇（Rhagoletis completaCresson：实蝇科）中分离出来的成团肠杆菌和肺炎克雷伯菌，并且博来霉素抗性可用作转化质粒的选择标记。我们转化了最初从与 R.completagut 密切相关的 Ent.agglomerans 和 K.pneumoniaes 菌株中产生增强型绿色荧光蛋白，这是绿色荧光蛋白的一种变体，首次证明了从实蝇害虫中分离出的内部胞外细菌的遗传转化。我们报告了这些细菌的质粒介导转化方法、转化质粒荧光标记蛋白的表达以及转化质粒在细菌中的稳定性。我们还讨论了该技术在害虫生物学研究和控制实施中的应用。
We discovered Zeocin™ is an effective antibiotic againstEnterobacter agglomeransandKlebsiella pneumoniaestrains isolated from the walnut husk fly (Rhagoletis completaCresson: Family Tephritidae) and that bleomycin resistance can be used as a selective marker in transforming plasmids. We transformedEnt.agglomeransandK.pneumoniaestrains originally isolated from their close association withR.completagut to produce enhanced green fluorescent protein, a variant of green fluorescent protein in the first demonstration of genetic transformation of internal extracellular bacteria isolated from a tephritid pest. We report methods for plasmid-mediated transformation of these bacteria, the expression of fluorescent marker protein from the transforming plasmids, and the stability of the transforming plasmid in the bacteria. We also discuss applications of this technology in the study of pest biology and control implementation.