Molecular Characterization and in Silico Analysis of a Novel Mutation in TEM-1 Beta-Lactamase Gene among Pathogenic E. coli infecting a Sudanese Patient

Hisham N Altayb^1, , Nagwa M El Amin², Maowia M. Mukhtar¹, Mohamed Ahmed Salih³ and Mohamed A M Siddig⁴

¹Institute of Endemic Diseases, University of Khartoum, Khartoum, Sudan

²Department of microbiology, Faculty of medicine, university of Khartoum, Sudan

³Head department of biotechnology, Biotechnology Park, Africa city of technology, Sudan

⁴Botany department, Faculty of Science, University of Khartoum, Sudan

Cite this paper:
Hisham N Altayb, Nagwa M El Amin, Maowia M. Mukhtar, Mohamed Ahmed Salih and Mohamed A M Siddig. Molecular Characterization and in Silico Analysis of a Novel Mutation in TEM-1 Beta-Lactamase Gene among Pathogenic E. coli infecting a Sudanese Patient. American Journal of Microbiological Research. 2014; 2(6):217-223. doi: 10.12691/ajmr-2-6-8

Abstract

The presence of ESBLs in many E. coli strains are of serious concern, since these organisms are the most common cause of different human infections. In this study we isolate an E. coli bacterium with high hydrolytic activity against cefotaxime. The ESBLs production was confirmed by phenotypic confirmatory test, while the ESBLs genes were detected by polymerase chain reaction (PCR). This isolate was positive for TEM gene and negative for CTX-M and SHV genes. DNA sequencing was done for TEM gene. The nucleotide sequences and translated proteins were subjected to BLAST for sequences similarity and homology, BLASTp result revealed a substitution of aspartic acid in TEM-1(gb: AFI61435.1) to Threonine at position 262. In Silico tools was used for mutation analysis and prediction of secondary and tertiary structure of wild and mutant type genes. We conclude that our mutant gene is completely different from the wild types TEM-1 gene, within phenotypic and genotypic levels. So we conclude a novel mutant TEM gene with ESBLs activity is been detected in Sudan Phylogenetic tree revealed that the possible source of our gene is Iran.

Keywords:
E. coli ESBLs Novel TEM gene Insilco analysis Sudan

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