| Tuberculosis (TB) is a chronic respiratory infectious disease caused by Mycobacterium tuberculosis (MTB). The only available vaccine against TB BCG has been extensively evaluated and demonstrated a variable protective efficacy ranging from 0 to 80% in different field trials. Furthermore, due to following issues, such as the problem of TB multidrug-resistant (MDR) strains,co-infection with HIV,and increasing mobility of population, the word-wide situation of TB was deteriorating,which has created an urgent need for new vaccines to prevent TB.After the human being infected, most the MTBs were licked up by macrophage , the others were mainly lived in macrophages, and the apoptosis of macrophages can kill the MTB to prevent them disseminating in vivo, therefore, the apoptosis ability of macrophage is very important for the fate of MTB. Because the macrophages expressed higher level of restrain apoptosis protein Mcl-1L, therefore, by inhibiting the expression of Mcl-1L which prompted macrophages apoptosis may be cure for tuberculosis.This study used siRNA technology to inhibit the expression of negative adjustment factor Mcl-1L The psimcl-1-1 and psimcl-1-2 as two expression plasmids of siRNA Targeting mcl-1L gene have been constructed, which means the purpose of double-stranded DNA coding siRNA has been cloned into siRNA expression vector psiRNA-hH1neo plasmid. After recombinant plasmids were transfected associated cells, the inhibition effects have been evaluated through real time PCR and Western Bolt analysis. Comparing the inhibitory effect, better siRNAmcl-1-2 has been choosed,and cloned with the anogher DNA fragment coding two TB common antigen into AAV vector as a novel type DNA vaccine integrated siRNA .This AAV vaccine joints two-factor interactions to enhance the protective efficacy to try to develop new tuberculosis DNA vaccine. The success of this approach would provide ideas and means for the prevention and therapy to other infectious diseases. |
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