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农杆菌介导的海胆胚胎转化

Agrobacterium-mediated transformation of sea urchin embryos.

作者信息

Bulgakov Victor P, Kiselev Konstantin V, Yakovlev Konstantin V, Zhuravlev Yuri N, Gontcharov Andrey A, Odintsova Nelly A

机构信息

Bioengineering Group, Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, Vladivostok, Russia.

出版信息

Biotechnol J. 2006 Apr;1(4):454-61. doi: 10.1002/biot.200500045.

Abstract

Agrobacterium-mediated transformation of higher plants is a well-known and powerful tool for transgene delivery to plant cells. In the present work, we studied whether Agrobacterium can transfer genetic information to animal (sea urchin) embryos. Sea urchin embryos were co-cultivated with A. tumefaciens strains carrying binary vectors containing the nptII marker gene and agrobacterial rolC and rolB oncogenes. Bacterial plasmid T-DNA-sea urchin DNA junction sites were identified in the genome of these embryos, thus indicating successful transformation. The nptII and both rol genes were expressed in the transformed embryos. The processes of transgene integration and transgene expression were suppressed when Agrobacteria contained mutated virA, virB or virG genes, suggesting that Agrobacterium transforms sea urchin cells by a mechanism similar to that which mediates T-DNA transfer to plants. Some of the embryos co-cultivated with Agrobacterium developed teratoma-like structures. The ability of Agrobacterium strains to trigger formation of teratoma-like structures was diminished when they contained the mutated vir genes. In summary, our results demonstrate that Agrobacterium is able to transform animal (sea urchin) embryonic cells, thus indicating a potential of this natural system for gene delivery to animal hosts. We also discuss the possibility of horizontal gene transfer from Agrobacterium to marine invertebrates.

摘要

农杆菌介导的高等植物转化是一种将转基因导入植物细胞的广为人知且强大的工具。在本研究中,我们研究了农杆菌是否能将遗传信息传递给动物(海胆)胚胎。海胆胚胎与携带含有nptII标记基因以及农杆菌rolC和rolB致癌基因的二元载体的根癌农杆菌菌株共同培养。在这些胚胎的基因组中鉴定出了细菌质粒T-DNA与海胆DNA的连接位点,从而表明转化成功。nptII基因和两个rol基因在转化后的胚胎中均有表达。当农杆菌含有突变的virA、virB或virG基因时,转基因整合和转基因表达过程受到抑制,这表明农杆菌通过一种类似于介导T-DNA转移到植物的机制来转化海胆细胞。一些与农杆菌共同培养的胚胎形成了畸胎瘤样结构。当农杆菌菌株含有突变的vir基因时,它们触发畸胎瘤样结构形成的能力减弱。总之,我们的结果表明农杆菌能够转化动物(海胆)胚胎细胞,从而表明这个天然系统具有向动物宿主进行基因传递的潜力。我们还讨论了从农杆菌向海洋无脊椎动物进行水平基因转移的可能性。

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