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Sadeghi D (msc), Mosavari N (phd), Rafiee B (msc), Mohamad Taheri M (msc), Dashtipour Sh (bsc), Zare A (phd), Ghahremanlo E (msc), Tebyanian M (phd),
Volume 6, Issue 1 (4-2012)
Abstract

Abstract Background and objectives: Tuberculin is the proteins existed in tuberculosis culture medium which precipitated by trichloroacetic acid (TCA) or ammonium sulfate. Tuberculin is used for diagnosis of Tuberculosis. The aim of this study is to compare the human tuberculin produced by Razi Institute and Mycobacterium tuberculosis Culture Filtrate Protein. Material and Methods: Initially By biphasic medium, Bacteria from Lowenstein–Jensen solid medium was transferred to a Dorset−Henley Liquid medium. After 6 weeks of growth, the bacteria were isolated from liquid medium containing secretory proteins by the 0, 22 micron filter and the solution containing secretory proteins was precipitated by TCA and ammonium sulfate, separately. Then, using spectrophotometer and kjeldahl protein assay, the presence of protein in solution was confirmed. At the end, the precipitated proteins are compared with the human tuberculin by Coomassie-Blue stained SDS-PAGE Results: The protein samples precipitated by TCA have more bands in the limit of higher than 20 kDa, but the protein samples by ammonium sulfate have more bands in the limit of less than 20 kDa. Human tuberculin proteins are like smear and their weight is less than 16 kDa. Conclusion: It seems that ammonium sulfate is more suitable for low molecular weight proteins than TCA for precipitation. Key words: Mycobacterium tuberculosis, SDS-PAGE, tuberculin
A Nazemi, N Vaseghi, M Khatamineja, A Nasrollahi Omran, M Eskandari,
Volume 7, Issue 5 (2-2014)
Abstract

Abstract Background and Objective: Recognizing and using of isolated phytase in the soil microorganisms are paramount importance to produce the Phytase enzyme utilized commercially in different industries. This study was conducted to recognize different bacillus species which are Phytase producers and detection of the gene that can produce this enzyme. Material and Methods: Soil samples were gathered through different parts of mountainous areas. The early isolation of bacillus was carried out in Bacillus Medium Agar. After isolating the bacteria and genome extraction, the responsible gene of enzyme producer recognized and amplified by PCR method. The size of this protein and the optimal production situation in supplemental exploitation such as SDS-PAGE and the enzymatic activity of its size were evaluated. Results: Of 40 samples, one bacterium secreting Phytase enzyme was isolated. This bacterium was sequenced and recognized Bacillus Sobtlis species that is classified in STR Genus. The size of protein phytase produced by this gene was about 45 KD and the enzyme activity at 55 degrees was measured about 5.65 in wavelength of 415 NM. The phytase gene with the size of 1200 bp was propagated. Conclusion: the microorganisms, in natural conditions, produce Phytase enzyme in limited amount and with the quality appropriate to microorganisms. Thus, isolating the bacilli producing Phytase enzyme and purifying this protein are highly significant. Key words: Bacillus Subtilis Phytase SDS-PAGE Enzymatic Activity Polymerization Chain Reaction
M Rasouli Nasab, Sh Habibnia, P Heidarieh, Mr Pourmand, M Fatahi, Ss Eshraghi,
Volume 7, Issue 5 (2-2014)
Abstract

Abstract Background and Objective: The Isolation of Nocardia species is complex and time-consuming, which is due to rapid growth of adjacent bacteria. Because of the importance of a specific medium with the ability of controlling intrusive microorganisms, this study aimed at comparing three laboratory methods to introduce the reliable isolation technique for Nocardia species. Material and Methods: The soil samples were collected from different regions of Tehran province, Iran, and carefully transferred to the laboratory. The samples were cultured in three different media including Paraffin Baiting,Humic acid vitamin B agar and Paraffin agar, and incubated for 3-4 weeks at 35 °C. Results: Of 110 soil samples, 31 Nocardia isolates (28.18%) were obtained from the media including Paraffin Baiting, (19 17.27%), Humic acid and vitamin B agar (4 3.63%), and Paraffin agar, (8 7.27%). Conclusion: because of high rate of isolation, low cost and the clearance of colonies suspected nocardia, Paraffin Bait technique is more reliable and efficient compared to the other methods. Key words: Nocardia Soil Paraffin Baiting Humic Acid Vitamin B

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