Construction of Genetic Recombination for Sea Cucumber Lysozyme in Bacillus subtilis

Abstract

Abstract: The expression of lysozyme from sea cucumber Stichopus japonicus in Bacillus subtilis were constructed by the method of recombinant DNA technique, and the expression plasmid pHT43-SjLys of growth curve and stability were analyzed. Results showed that a specific strip about 400 bp was visible in the result of agarose gel electrophoresis, which was consistent with expected size of sea cucumber lysozyme gene. Construction of the lysozyme was performed in a vector pHT43 and validated by double digestion with BamHⅠand SmaⅠ, witch indicated the fragment size was consistent with the expected result. The growth trend of engineering bacteria was consistent compared with the wild type strain WB600, and insertion of foreign genes did not affect the cell’s metabolism. In the absence of selection pressure, the recombinant plasmid was stability, and there was no gene rearrangement and lost. The results showed that the recombinant sea cucumber lysozyme genetic engineering bacteria pHT43-SjLys/WB600 is constructed successfully.

Key words: Genetic recombination, Sea cucumber lysozyme, Bacillus subtilis, Genetic engineering bacteria, Expression vector

Sun Lu, Liu Zhiwen, Zou Dan, Li Dan, Pan Bo, Cong Lina. Construction of Genetic Recombination for Sea Cucumber Lysozyme in Bacillus subtilis[J]. Biotechnology Bulletin, 2014, 0(6): 150-154.

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