Department of Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT, UK.
Synthace Ltd, The Westworks, 195 Wood Lane, London, W12 7FQ, UK.
Biotechniques. 2020 Nov;69(5):384-387. doi: 10.2144/btn-2020-0101. Epub 2020 Nov 2.
Here we present a modification of the widely used pET29 expression vector for use in rapid and straightforward parallel cloning via a gene replacement and Golden Gate strategy. The modification can be applied to other expression vectors for Gram-negative bacteria. We have used the modified vectors to clone large numbers of bacterial natural enzyme variants from genomic and metagenomic sources for applications in biocatalysis.
在这里，我们介绍了对广泛使用的 pET29 表达载体的一种修改，用于通过基因替换和 Golden Gate 策略进行快速而直接的并行克隆。该修改可应用于革兰氏阴性细菌的其他表达载体。我们已经使用修饰的载体从基因组和宏基因组来源克隆了大量的细菌天然酶变体，用于生物催化应用。
