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Author:
Date:
2014-03-05
[Abstract] The genome-wide screen Tn-seq (van Opijnen et al., 2009) is very valuable tools to identify bacterial genes with a conditionally essential function, for instance genes involved in bacterial virulence. These techniques are based on the generation of a random mutant library, which is grown in a control of challenge situation (Figure 1). The advantage of using a mariner transposon for the generation of a random transposon mutant library is its insertion into TA sites, which makes the insertion in the genome highly random. In addition, an MmeI restriction site can be introduced in the inverted repeat of the transposon, without affecting the recognition by HimarC9 transposase.
[摘要] 全基因组筛选Tn-seq（van Opijnen等人，2009）是鉴定具有条件必需功能的细菌基因（例如涉及细菌毒力的基因）的非常有价值的工具。 这些技术基于产生随机突变体文库，其在挑战情况的对照中生长（图1）。 使用水手转座子产生随机转座子突变体文库的优点是其插入TA位点，这使得在基因组中的插入高度随机。 此外，可以在转座子的反向重复中引入MmeI限制位点，而不影响HimarC9转座酶的识别。
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