Construction expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human human B7-2
Abstract：Objective To construct a recombinant bacillus Calmette-Gutrin vaccine (rBCG) secreting hu-man B7-2. Methods The DNA fragment encoding human hB7-2 was amplified from the plasmid containing hB7-2 by using polymerase chain reaction (PCR) and then inserted into the shuttle expression vector pYL-MCS. The re-combinant plamid pYL-hB7-2 was identified by restriction endonuclease digestion, PCR amplification and nucleotide sequencing, pYL-hB7-2 was electroperated into BCG to get rBCG. The DNA of hB7-2 gene in rBCG were deter-mined by PCR and nucleotide sequencing. The protein expression of hB7-2 was detected by SDS-PAGE and enzyme-linked immunosorbent assay (ELISA) respectively. Results The hB7-2 sequence derived from rBCG by PCR was the same as that reported previously. The recombinant plamid pYL-hB7-2 was constructed successfully and con-firmed by restriction endonuclease analysis, PCR detection and nueleotide sequencing analysis, pYL-hB7-2 was suc-cessfully transformed into BCG by electroporation and was capable of synthesizing and secreting hB7-2 by SDS-PAGE. The level of hB7-2 in the culture supernatant of rBCG was 3.8U/ml. Conclusions The constructed recom-binant BCG strain produces and secretes costimulator human B7-2, which will provide a laboratory foundation for po-tential clinical use for bladder cancer immunotherapy.
Keywords：Bladder tumorBacillus calmette guerinCostimulatorGene recombinant
Publication Date：2009-01-01
Online Publishing Date：2026-09-14(First online date of this platform, not the publication date of the document)
Pages：3( 944-946 )