ISOLASI DAN KARAKTERISASI FRAGMEN 0,9 KB DNA LOKUS PYR A (IVIL) SALMONELLA TYPHIIMURIUM

• Main Author: Nafisah, Evi
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/4672
• Institution: Institut Teknologi Bandung

<b>Abstract:<p align=\"justify\"> <br /> <br /> A study of genes that are responsible to Salmonella typhi pathogenesis is needed to understand the etiology of typhoid. Mekalanos group developed the IVET (In vivo Expression Technology) and discovered five genes which are thought to be responsible for Salmonella typhimurium pathogenesis in mice so that it has served as a model to investigate the pathogenecity of S. typhi. One of the five genes that was discovered by Mekalanos is ivil, the gene that has homology with the carAB operon of E. coli K-12. The general aim of this research is to study the molecular mechanism of S. typhi pathogenecity. Particularly, this research was planned to isolate and characterise a S typhimurium 0.9 kb DNA fragment. A set of CA-3/CA-6 primers that was designed based on the carAB operon of E. coil K-12 amplified the 0.9 kb S. typhimurium DNA fragment succesfully. The positive 0.9 kb PCR product then was cloned using T-vector pMOSB1ue in E. coil MOSBIue. The resulting clones were confirmed by sequencing through dideoxy Sanger method. The sequencing result showed that the 0.9 kb DNA fragment contained 903 bp that consisted of 676 bp partial carA gene, 15 bp intercistronic region, and 212 bp partial carB gene of S. typhimurium. Analysis of nucleotide homologue between the 0.9 kb S. typhimurium DNA fragment and the car AB operon of S. typhimurium LT2 (GenBank, 1996) was 95%. Moreover, the nucleotide homologue between the 0.9 kb S. typhimurium DNA fragment with the carAB operon of E. coli K-12 and S. typhi respectively was 85% and 93 %. Further more analysis of amino acids homologue showed that there is 97% homologue between the 0.9 kb S. typhimurium DNA fragment and the carAB operon of S. typhimurium LT2 (GenBank, 1996). The amino acids homologue between the 0.9 kb S. typhimurium DNA fragment with the car AB operon of K coil K-12 and gen carAB of S. typhi were 93,5 % and 86 % respectively.<p align=\"justify\">