Preliminary Proteomic Study On The Multidrug Resistant Strain CCDC5180 And Drug Sensitive Strain CCDC5079 Of Beijing Family Of Mycobacterium Tuberculosis And BCG

Mycobacterium tuberculosis (MTB)is one of a serious pathogen affected human healthin which Beijing Family was the main genetic typing that causes prevalence of tuberculosis(TB) in China. Multidrug resistant strain CCDC5180 and Drug sensitive strain CCDC5079were two strains among Beijing family that were selected for genome sequencing. Thevaccine BCG was widly used to prevant and control TB, but the protection to adult is invalid.In order to investigate MTB's biologic characteristics and the mechanism of drugresistantce, proteomic of MTB was preliminarily conducted with CCDC5180, CCDC5079and BCG by means of sodium dodecul sulphonate denatured polyacrylamide gelelectrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE) and massspectrometry (MS).The SDS-PAGE results showed that bands of whole cell proteins in the three strains werehighly similar to each other. There were about 40 bands in the pattens of the whole cellproteins. The 14.8KD,23.6KD and 62.0KD were highly expressed and majority of proteinswere approximately located between the 25.0KD-84.2KD range, but it is difficult to clarifythe differences of strains. Therefore, the 2-DE was applied for getting clearer profile of thedifferences between 3 strains,commercially available 24cm IPG with pH4-7 was served asthe first dimension,and 12.5% polyacrylamide gel was served as second-dimension electrophoresis. The patterns of the three strains investigated were highly similar,approximately 1563,1620 and 1654 protein spots were observed on 2-DE gels ofCCDC5180,CCDC5079 and BCG respectively, depending in the condition of silver-stainingconditions, most protein spots were distributed in the range of PH4.0-6.5. silver-staining ishigh-resolution, but low identification of MS, so we must use 2-DE gels stained byCoomassie Brilliant Blue G-250 to identify by MALDI-TOF-M and MS/MS. We separatedthe whole cell protein of CCDC5079,CCDC5180 and BCG strains with 2-DE, stained byCoomassie Brilliant Blue G250, 718,767,1019 spots were observed, identified 443,436,486 protein spots information respectivly, finally acquired 226,149,165 difference sortsprotein. By bioinformatic analysising, we obtained the protein expression data including genename,related ORF,founctions and other basic informations. Some unidentified proteinsmaybe the specific ones of Beijng Family of MTB and need to further study carefully. The2-DE patterns differences between CCDC5079 and CCDC5180 show that, 7 protein spotsincreased intensity, 4 protein spots decreased intensity in CCDC5079, 5 were unique toCCDC5079 and 2 to CCDC5180. Between CCDC5079 and BCG, 5 protein spots increasedintensity, 8 protein spots decreased intensity in CCDC5079,7 were unique to CCDC5079 and2 to BCG. Compared the differences of CCDC5180 with BCG, 4 protein spots increasedintensity, 18 protein spots decreased intensity in CCDC5180, 2 were unique to CCDC5180and 4 to BCG.. By analysising the protein expression data and 2-DE patterns differences, wewill get more mechanism information of drug resistant, provide the theory for developing newvaccine and specific diagnosis reagent, and establish the fine base for proteomic study ofBeijing Family of MTB.