蚊子基因组的高效位点特异性基因工程。
High efficiency site-specific genetic engineering of the mosquito genome.
作者信息
Nimmo D D, Alphey L, Meredith J M, Eggleston P
机构信息
Centre for Applied Entomology and Parasitology, Keele University, Keele, Staffordshire UK.
出版信息
Insect Mol Biol. 2006 Apr;15(2):129-36. doi: 10.1111/j.1365-2583.2006.00615.x.
Abstract
Current techniques for the genetic engineering of insect genomes utilize transposable genetic elements, which are inefficient, have limited carrying capacity and give rise to position effects and insertional mutagenesis. As an alternative, we investigated two site-specific integration mechanisms in the yellow fever mosquito, Aedes aegypti. One was a modified CRE/lox system from phage P1 and the other a viral integrase system from Streptomyces phage phi C31. The modified CRE/lox system consistently failed to produce stable germline transformants but the phi C31 system was highly successful, increasing integration efficiency by up to 7.9-fold. The ability to efficiently target transgenes to specific chromosomal locations and the potential to integrate very large transgenes has broad applicability to research on many medically and economically important species.
摘要
目前用于昆虫基因组基因工程的技术利用了转座遗传元件，这些元件效率低下、承载能力有限，并会产生位置效应和插入诱变。作为替代方案，我们研究了黄热病蚊子埃及伊蚊中的两种位点特异性整合机制。一种是来自噬菌体P1的改良型CRE/lox系统，另一种是来自链霉菌噬菌体phi C31的病毒整合酶系统。改良型CRE/lox系统一直未能产生稳定的种系转化体，但phi C31系统非常成功，将整合效率提高了7.9倍。将转基因有效靶向特定染色体位置的能力以及整合非常大的转基因的潜力，在许多医学和经济上重要物种的研究中具有广泛的适用性。
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Plant Biotechnol J. 2004 Mar;2(2):169-79. doi: 10.1111/j.1467-7652.2003.00061.x.
2
Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster.通过黑腹果蝇中转座子载体末端序列缺失实现整合后稳定化
Nat Biotechnol. 2004 Sep;22(9):1150-4. doi: 10.1038/nbt1002. Epub 2004 Aug 8.
3
Construction of transgenic Drosophila by using the site-specific integrase from phage phiC31.利用来自噬菌体phiC31的位点特异性整合酶构建转基因果蝇。
Genetics. 2004 Apr;166(4):1775-82. doi: 10.1093/genetics/166.4.1775.
4
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5
Gene vector and transposable element behavior in mosquitoes.基因载体与转座元件在蚊子中的行为表现
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