| Autor: |
Xiao-qiang, Yang, Chun-xiu, Peng, Bo, Jiang, Rui, Xing, Shao-rong, Zhang, Xue-qing, Chen |
| Rok vydání: |
2005 |
| Předmět: |
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| Zdroj: |
Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA. 25(10) |
| ISSN: |
1000-2588 |
| Popis: |
To construct and characterize an exogenous gene expression system in Lactococcus lactis.PCR-based gene assembly was used to synthesize the gene sequence containing P59 promoter, USP45 signal peptide, ribosome binding site and multiple cloning site. Plasmid pBS-pu was obtained after ligation of the assembled sequence with pBluescript II SK (+), and the terminator of USP45 protein was added to construct the recombinant plasmid pNBC1000. The gene coding for Streptococcus pyogenes M6 protein was amplified and cloned into pNBC1000 to obtain the plasmid pNBC2000. To characterize the expression system, enhanced green fluorescent protein (EGFP) gene was amplified from the plasmid pEGFP-N1 and cloned into pNBC2000. Laser scanning confocal microscope was used to observe the bacteria containing pNBC2000, pNBC2000-EGFP or pBS-EGFP.Green fluorescence was visualized in the bacteria containing pNBC2000-EGFP, but not in the bacteria containing pNBC2000 or pBS-EGFP.The plasmids pNBC1000 and pNBC2000 containing P59 promoter, USP45 signal peptide, ribosome binding site and multiple cloning sites are successfully constructed, which are capable of expressing exogenous gene in Lactococcus lactis. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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