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发明公开
CN107502619A 一组干酪乳杆菌基因敲除载体及其应用 有权

一组干酪乳杆菌基因敲除载体及其应用
摘要：
本发明公开了一组干酪乳杆菌基因敲除载体，包括一个介导外源双链DNA敲除盒up-lox66-cat-lox71-down与染色体上同源序列之间的同源重组的专用载体pMSP456和一个介导lox66和lox71位点之间的重组反应以便切除抗性基因的专用载体pMSPcre；其中载体pMSP456的核苷酸序列如SEQ ID No.1所示，载体pMSPcre的核苷酸序列如SEQ ID No.2所示。本发明还公开了所述载体在构建干酪乳杆菌基因敲除突变株中的应用。实验证实利用本发明的载体pMSP456和pMSPcre敲除干酪乳杆菌基因，其敲除效率达到100％，且操作简单，耗时短，在干酪乳杆菌代谢工程中具有广阔的应用前景。
摘要（英）：

The invention discloses a Lactobacillus casei gene knockout vector group. The Lactobacillus casei gene knockout vector group comprises a special vector pMSP456 which mediates the homologous recombination between a foreign double-stranded DNA knockout box up-lox66-cat-lox71-down and a homologous sequence on a chromosome, and a special vector pMSPcre which mediates the recombination reaction between loci lox66 and lox71 in order to excise a resistant gene. The nucleotide sequence of the vector pMSP456 is shown in SEQ ID No. 1, and the nucleotide sequence of the vector pMSPcre is shown in SEQ ID No. 2. The invention further discloses the application of the vectors to construction of a Lactobacillus casei gene knockout mutant strain. Experiments show that by using the vectors pMSP456 and pMSPcre to knock out a Lactobacillus casei gene, knockout efficiency reaches 100%, operation is easy, time consumption is low, and the vectors have broad application prospects in Lactobacillus casei metabolic engineering.

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