Sequence analysis of endogenous plasmid of Lactobacillus plantarum and construction of its expression vector

Abstract

Abstract: （Objective）In recent years, more and more researches on genetic tools of lactic acid bacteria have been carried out. The sequence analysis and functional analysis of the plasmids carried by the strains have led to the construction of Lactobacillus expression vectors, which is the main direction to promote the development of Lactobacillus genetic engineering. （Methods）Plasmids were extracted from Lactobacillus plantarum LP3 and sequenced and functionally analyzed. An Escherichia coli/Lactobacillus shuttle vector was constructed based on the pLP3, the chloramphenicol resistance gene from pSCPSP as a selection marker and the replicon of pUC19. Furthermore, an expression vector D-pLP3-PslpA was developed with the promoter of the promoter of S-layer protein SlpA from Lactobacillus acidophilus. The green fluorescent protein(GFP) as a reporter was expressed in Lactobacillus plantarum tested. （Results）a cryptic plasmid pLP3 was obtained from lactobacillus plantarum LP3 which isolated from traditional fermented sauerkraut. The size of pLP3 was 2017bp and GC content was 37.48%. Based on the backbone of pLP3,we constructed an Escherichia coli / Lactic acid bacteria shuttle vector D-pLP3 and electroporated it into several strains of Lactic acid bacteria. The transformation efficiencies were ranged from 0.3 * 10 ^ 2 ~ 1.0 * 10 ^ 3 cfu/ug . The green fluorescent protein(GFP) as a reporter was expressed successfully in Lactobacillus plantarum tested, (Conclusion) These results indicated that the expression vectors D-pLP3-PslpA possesses the potential to be used as a molecular tool for heterologous gene cloning and expression in lactobacilli.

Key words: cryptic plasmid, Rolling-circle-replication, chuttle vector, Lactobacillus plantarum, Construction of lactic acid bacteria expression vector

CLC Number:

FangLai-shan. Sequence analysis of endogenous plasmid of Lactobacillus plantarum and construction of its expression vector[J]. FOOD SCIENCE, 0, (): 0-0.

References

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