Cloning And Expression Of Alkaline Protease Gene From Bacillus Stearothermophilus Strain Fl
Ab.Hamid, Suhaili (2000) Cloning And Expression Of Alkaline Protease Gene From Bacillus Stearothermophilus Strain Fl. Masters thesis, Universiti Putra Malaysia.
Cloning of alkaline protease gene from a thermophilic bacteria, Bacillus stearothermophilus Fl (BSF1) was done by PCR cloning method. Genomic DNA of BSFI was extracted and the highest concentration obtained was 0.46 ug/uL and with a purity of 2.0. Three sets of primers were used including RM5 (5'-CA(CT) GG(ACGT) ACC AA(CT) GTG GC(CGT) GG-3) and RM6 (5'-(ACG)GG GGT (ACG)GC CAT GGA (CGT)CC-3', F0R900 (5-GCA TGC TAC GAT TAA ATA TC-3) and REV900 (5'-CGG CAA TAT CAC ITA GAG TAC C-3') and FOR900 (5'-GCA TGC TAC GAT TAA ATA TC-3') and REV1 591 (5-TGC AGC AGA AAG AAG GAA-3') which resuhed in amplification of 500-bp, 900-bp and 1 500- bp products, respectively. Southern blotting analysis suggested that both 500-bp and 900-bp fragment were present within the 1 500-bp fragment. E. coli TOPIOF
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