STUDI KEANEKARAGAMAN NUKLEOTIDA GEN CARA BAKTERI DAN MANUSIA MENGGUNAKAN TEKNOLOGI PCR

• Main Author: Ihsanawati
• Format: Theses
• Language: Indonesia
• Subjects: Biokimia
• Online Access: https://digilib.itb.ac.id/gdl/view/4686
• Institution: Institut Teknologi Bandung

<b>Abstract:<p align="justify"> <br /> <br /> The carAB operon of E. coil K-12 encodes two subunits of the carbamoyl phosphate sintetase. This enzyme plays an important role for arginine and pyrimidine biosynthesis in all organisms. The carAB operon therefore is referred to as a housekeeping gene. Despite the complete-obtained nucleotide sequence of the carA genes of E. colt K-12 and S. typhi Lister, very little is known about other bacterial carA and carB genes. PCR can be employed to detect nucleotide diversity among bacterial carA genes using primers that were designed based on the carA genes of both of bacteria. The main objective of this research was to study the nucleotide diversity among bacterial and human carA genes through PCR technology. In order to achieve this goal, 18 Salmonella sp., 22 non-Salmonella sp., and human DNAs were amplified by PCR using three set of CA-1/CA-2, CA-7/CA-8, and CA-8/CA-9 primers. All DNAs that were amplified with the CA-1/CA-2 primers showed the positive 0.6 kb PCR product. The template DNAs from 14 of 40 bacteria including DNA of S. typhi resulted the 0.4 kb DNA fragment with the CA-7/CA-8 primers whereas only S. typhi was amplified employing the CA-8/CA-9 primers. All PCR products with the CA-1/CA-2, the CA-7/CA-8, and the CA.-8/CA-9 primers revealed nucleotide diversity among Salmonella sp., non-Salmonella sp., and human carA genes. In addition, the CA-8/CA-9 primers were highly specific for S. typhi. These findings suggested that the PCR assay based on the unique sequence in the carA gene of S. typhi could be used to develop a rapid and sensitive diagnostic test for typhoid fever.<p align="justify">