Original Articles
Armaini, Abdl Dharma and Marni
Abstract
Isolates of 1-M3 isolated from a hot spring Rimbo Panti, has been identified using 16S rRNA gene. Based on the results of BLAST and kinship analysis, found that isolates RP 1-M3 including Bacillus sp. Amplification with primers specific for the gene endo-1,4 β-Glucanases obtained ± 1500 bp PCR product. PCR products were cloned into plasmid pGEM-T Vector obtained recombinant plasmid. Recombinant plasmid transformation into E. coli host, and selection results of transformation using media containing antibiotics, IPTG and X-GAL to get the positive transformation that carries a recombinant plasmid with target DNA. To observe the success of the transformation carried out colony PCR with primers T7SP6, obtained ± 2.000 bp PCR product consisting of 1,500 bp, indicated as DNA gene coding the enzyme endo-β1,4-Glucanases and ± 500 bp of sequence T7SP6. A result of gene sequence was translated into amino acids obtained Egl protein of 500 amino acid sequence. Based on the results of BLAST in the GenBank data base, the genes that have been isolated from Bacillus sp. RP 1-M3 includes genes endo-β1,4- Glucanases with 99.4% similarity with genes endo-β1,4-Glucanases from Bacillus megaterium (AGW99981.1) at the amino acid level.