Sleight Sean C, Sauro Herbert M
Department of Bioengineering, University of Washington, Seattle, WA, USA.
Methods Mol Biol. 2013;1073:19-30. doi: 10.1007/978-1-62703-625-2_3.
Synthetic biologists assemble genetic circuits from standardized biological parts called BioBricks™. BioBrick™ examples include promoters, ribosome binding sites, DNA or RNA-coding sequences, and transcriptional terminators. Standard BioBrick™ assembly normally involves assembly of two BioBricks™ at a time using restriction enzymes and DNA ligase. Here we describe an alternative BioBrick™ assembly protocol that describes the assembly of two BioBricks™ using the In-Fusion PCR Cloning Kit. This protocol can also be adapted to use similar recombination-based assembly methods, such as SLIC and Gibson assembly.
合成生物学家利用称为BioBricks™的标准化生物元件组装遗传电路。BioBrick™的例子包括启动子、核糖体结合位点、DNA或RNA编码序列以及转录终止子。标准的BioBrick™组装通常一次使用限制性内切酶和DNA连接酶组装两个BioBrick™。在此，我们描述了一种替代的BioBrick™组装方案，该方案描述了使用In-Fusion PCR克隆试剂盒组装两个BioBrick™。该方案也可适用于使用类似的基于重组的组装方法，如SLIC和吉布森组装。
