Part:BBa_K2556051
f1-AraC-Pbad-lysis-f2
随着重组DNA技术得到广泛应用以来，生物技术发生了质的飞跃。很多基因工程产物都是胞内物质，必须将细胞破壁，使产物得以释放，才能进一步提取，因此细胞破碎是提取胞内产物的关键步骤，破碎方法的得当与否，直接影响到所提取产品的产量、质量和生产成本。近年来常用的几种细胞破碎方法有以下几种：高压匀浆法、高速珠磨法、超声破碎、酶溶法以及化学渗透法。这些方法或存在损害产物、或是成本高或是效率低的问题，为此我们项目拟构建一株加入阿拉伯糖后可以自行裂解的工程菌。我们先在来自噬菌体的裂解基因前添加阿拉伯糖操纵子，再通过CRISPR/Cas9技术，将AraC-Ara Promoter-lysis元件构建到E.coli MG1655野生株的基因组中。 With the wide application of recombinant DNA technology, biotechnology has undergone a qualitative leap. Many of the genetic engineering products are intracellular substances, so the cell wall must be broken down so that the product can be released before it can be further extracted, so cell fragmentation is a key step in extracting intracellular products, and the method of fragmentation is appropriate or not. Directly affect the output, quality and production cost of the extracted products. In recent years, the methods of cell fragmentation are as follows: high pressure homogenization, high speed bead milling, ultrasonic crushing, enzyme dissolution and chemical permeation. These methods may have harmful products, high cost or low efficiency. For this reason, we plan to construct a strain of engineering bacteria that can be cracked by adding arabinose. We put AraC-Ara Promoter in front of the lytic gene from phage, and then we use CRISPR/Cas9 technology to turn the construction of AraC-Ara Promoter-Lysis into the genome of E.coli MG1655 wild strain. This part consists of fragments from the E.coli MG1655 genome, AraC-AraPromoter and lytic genes from phages.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2200
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2826
Illegal NgoMIV site found at 3636
Illegal AgeI site found at 2035
Illegal AgeI site found at 4001 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 2017
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