Recombinant lactococcus lactis for secreting and expressing Dermatophagoides pteronyssinus allergen Derp2 and application of recombinant lactococcus lactis
A technology for secreting and expressing Lactococcus lactis, which is applied to the recombinant Lactococcus lactis that secretes and expresses the house dust mite allergen Derp2 and its application field, which can solve the problems of low purity, hindering the clinical application of dust mite allergen reagents, and complicated operations
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[0026] The present invention will be described in more detail below in conjunction with specific embodiments.
[0027] 1. According to the genome data of Lactococcus lactis, analyze the parameters such as codon preference and GC content of Lactococcus lactis, the natural gene sequence of the main allergen Derp2 derived from the house dust mite (Dermatophagoides pteronyssinus) The modified allergen gene The sequence is SEQ ID No.2 (genebank: AF276239.1) for codon optimization, and the gene mderp2 that can be highly expressed in Lactococcus lactis is obtained, and the sequence of the gene is SEQ ID No.1;
[0028] 2. The optimized gene mderp2 was cloned into the expression plasmid pNZ8148 of Lactococcus lactis (gifted by Professor Zhong Jin of the Chinese Academy of Sciences) to obtain the secretory expression plasmid pNZ8148-SD, and the expression plasmid was electrotransformed into the host strain L. lactis NZ9000 ( The recombinant strain L.lactis SD was obtained from the Chine...
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