Magnetic capture-hybridization method for purification and probing of mRNA for neutral protease of Bacillus cereus.
نویسندگان
چکیده
A magnetic capture-hybridization method was assessed for the isolation of prokaryotic mRNA for the neutral protease of B. cereus from liquid culture. A biotin-labeled specific probe was hybridized to the mRNA transcripts and subsequently captured by streptavidin-coated paramagnetic beads. mRNA was detected by dot-blot hybridization with a ds DIG-labeled DNA-probe. The magnetic capture hybridization is a rapid and simple method and has a promising potential for gene expression studies in complex samples.
منابع مشابه
Specific detection of the gene for the extracellular neutral protease of Bacillus cereus by PCR and blot hybridization.
A pair of primers and a gene probe for the amplification and detection of the Bacillus cereus neutral protease gene (NPRC) were developed. Specificity for the npr genes of the B. cereus group members B. cereus, B. mycoides, and B. thuringiensis was shown. Restriction polymorphism patterns of the PCR products confirmed the presence of the NPRC gene in all three species.
متن کاملPurification and Properties of a Collagenolytic Protease Produced by Bacillus cereus MBL13 Strain
A novel collagenase-producing bacterium has been isolated and identified as Bacillus cereus MBL13. From the culture supernatant of B. cereus MBL13 grown on bone collagen as the sole carbon and nitrogen source, an extracellular protease with novel property of hydrolyzing waste animal bones was purified. The molecular mass of the purified collagenolytic protease was estimated to be (38.0±1.5) kDa...
متن کاملPurification and characterization of laundry detergent compatible alkaline protease from Bacillus cereus
An alkalophilic bacterium, Bacillus cereus produced an extracellular alkaline protease, which was found to be active at high temperature and pH range, suitable for commercial laundry detergents. B. cereus protease was partitioned in different aqueous two-phase systems such as PEG/dextran, PEG/potassium phosphate, PEG/sodium citrate, PEG/magnesium sulphate and PEG/sodium dihydrogen phosphate and...
متن کاملIsolation, Purification and Characterization of a Thermophilic Alkaline Protease from Bacillus subtilis BP-36
The goal of this research was to isolate and identify the thermostable alkaline protease producing bacteria among several native Iranian microorganisms. At the end of screening program, a Bacillus subtilis BP-36 strain producing thermophilic alkaline protease was isolated from a hot spring in Ardebil province. The target enzyme was purified using a one-step Aqueous two-phase systems (ATPS) prot...
متن کاملCharacterization of an a-Amylase with Broad Temperature Activity from an Acid-Neutralizing Bacillus cereus Strain
Bacillus sp. GUF8, isolated from acidic soil samples of a tea farm was identified as Bacillus cereus, based on 16S rDNA sequencing and standard bacterial identification methods. Following optimization of enzyme production, the resulting α-amylase was purified by acetone precipitation and ion exchange chromatography. Consequently, thermostability and kinetic parameters of the purified enzyme wer...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Journal of microbiological methods
دوره 37 2 شماره
صفحات -
تاریخ انتشار 1999