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钩端螺旋体LipL21基因真核表达载体pcDNA3.1(+)/LipL21的构建与鉴定

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TL;DRAbstract

Objective To construct the eukaryoctic expression plasmid pcDNA3.1(+)/LipL21 containing the outer membrane Lipoprotein LipL21 of the Leptospira interrogans serovar Lai, and provide a new candidate antigen for preventing leptospirosis. Methods LipL21 Gene was amplified from the genomic DNA of Leptospira interrogans serovar Lai, strain Lai 56601 by polymerase chain reaction (PCR), and the gene was inserted into cloning vector pUCm-T. The inserted LipL21 gene was subcloned into appropriate site of pcDNA3.1(+) vector. The positive clones were screened after being identifid with restrictive enzymes and sequence analysis. After being sequenced with DNA auto-sequence analysis instrument, the amplified DNA sequence of LipL21 was searched for alignment with NCBI Blast program. Results The target gene LipL21 segment about 561bp was obtained successfully. Homology of the nucleotide and putative amino acid sequence of LipL21 gene between Leptospira interrogans serovar Lai, strain Lai 56601 was 100

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Objective To construct the eukaryoctic expression plasmid pcDNA3.1(+)/LipL21 containing the outer membrane Lipoprotein LipL21 of the Leptospira interrogans serovar Lai, and provide a new candidate antigen for preventing leptospirosis. Methods LipL21 Gene was amplified from the genomic DNA of Leptospira interrogans serovar Lai, strain Lai 56601 by polymerase chain reaction (PCR), and the gene was inserted into cloning vector pUCm-T. The inserted LipL21 gene was subcloned into appropriate site of pcDNA3.1(+) vector. The positive clones were screened after being identifid with restrictive enzymes and sequence analysis. After being sequenced with DNA auto-sequence analysis instrument, the amplified DNA sequence of LipL21 was searched for alignment with NCBI Blast program. Results The target gene LipL21 segment about 561bp was obtained successfully. Homology of the nucleotide and putative amino acid sequence of LipL21 gene between Leptospira interrogans serovar Lai, strain Lai 56601 was 100

Keywords

Leptospira interrogansGeneBiologyLeptospiraPlasmidgenomic DNANucleic acid sequenceRecombinant DNA

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