Intramolecular integration assay validates integrase phi C31 and R4 potential in a variety of insect cells.

Jakkrawarn Chompoosri, Tresa Fraser, Yupha Rongsriyam, Narumon Komalamisra, Padet Siriyasatien, Usavadee Thavara, Apiwat Tawatsin, Malcolm J Fraser
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Abstract

Phage phi C31 and R4 integrases are site-specific and unidirectional serine recombinases. We have analyzed the ability of these integrases to mediate intramolecular integration between their attB and attP sites in 7 important insect cell lines as a means of predicting their relative mobility in the corresponding insect species. Both integrases exhibit significantly higher frequencies in Drosophila S2 cells than in the other insect cell lines examined, but do work well in all of the species tested. Our results, coupled with previous results of the activity of phi C31 integrase in D. melanogaster and Aedes aegypti, suggest the family of serine catalyzed integrases will be useful site-specific integration tools for functional genome analysis and genetic engineering in a wide range of insect species.

分子内整合实验验证了整合酶phi C31和R4在多种昆虫细胞中的潜力。
噬菌体phi C31和R4整合酶是位点特异性和单向丝氨酸重组酶。我们分析了这些整合酶在7种重要昆虫细胞系中介导attB和attP位点分子内整合的能力,以此预测它们在相应昆虫物种中的相对迁移能力。这两种整合酶在果蝇S2细胞中比在其他昆虫细胞系中表现出更高的频率,但在所有被测试的物种中都表现良好。我们的研究结果,再加上之前在黑脉伊蚊和埃及伊蚊中phi C31整合酶的活性结果,表明丝氨酸催化整合酶家族将成为广泛昆虫物种功能基因组分析和基因工程中有用的位点特异性整合工具。
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