Objective To obtain highly expressing cell lines by inserting the glutamine synthetase (GS) screening system and replacing the promoter of the vector.Methods The mutation of the point BamHⅠwas induced to build a new vector pIRES2-EGFP.The marker gene GS was inserted by AseⅠ and NheⅠ, and the promoter hCMV was replaced by PacⅠand NheⅠ.The new vector pHGS1.0 and the vector pIRES2-enhanced screen fluorescein protein( EGFP)-B were inserted by the recombinant protein TEM8 ( 1-227 )-VEGFR1 domain2-IgG2 ( TV-IgG2 ) gene to analyze the advantages of the expression.Results The glutamine synsthetase is successfully inserted, the human cytomegalovirus replaced, and recombinant protein is increased 5-fold by human immunoglobulin quantification kit.Conclusion The GS system is a highly protein expressing system.

Objective To investigate the immune protection of heparin-hinding hemagglutinin ad-hesin(HBHA) and to estimate its potential diagnostic value. Methods Native HBHA were used to stimu-late peripheral blood mononuelear cells (PBMCs) from different infected-cases including PPD negative healthy control, PPD positive latent tuberculosis(LTB) infection, pulmonary tuberculosis, and the IFN-γ/in the supernatant of culture was detected. Meanwhile, HBHA specific IgG antibody in the sera was detected by ELISA. Results The middle level of HBHA specific IFN-γ of the three groups were 49.5 pg/ml, 781.9 pg/ml and 341.8 pg/ml, respectively. IFN-γ of latent tuberculosis group was much higher than that of the control, and slightly higher than that of the patients with pulmonary tuberculosis. And the absorbency of the IgG antibody to HBHA in the three groups was 0.212±0.066, 0.224 ± 0.076 and 0.285±0.078. lgG an-tibody in the patients with pulmonary tuberculosis is higher than that of the healthy, including the control and the latent tuberculosis infection. Conclusion HBHA has good immunogenieity, and it can stimulate the LTB to release high level IFN-γ, suggests that the LTB doesn't develop active tuberculosis may rely on its protection. HBHA specific. IFN-γ release may identify 1,333 from the healthy. Anti-HBHA antibody plays an auxiliary role in the diagnosis of pulmonary tuberculosis.