الكشف الجزيئي عن بكتريا Oxalobacter formigenes باستخدام جينات 16SrRNA، oxc في البراز من اشخاص اصحاء == Molecular Identification of Oxalobacter Formigenes Bacteria Based On 16S rRNA, Oxc Genes In Stool Samples Healthy Subjects
Author name:
طارق طلال خلف
Supervisor name:
زهرة محمود الخفاجي
General topic:
Biology
Specific topic:
Biotechnologies
Degree:
Master
University:
University of Baghdad
Language:
Arabic
University location:
Baghdad
First pages:
24T2781 - p.pdf
Abstract:
بكتريا Oxalobacter formigenes من مكونات المكنون البشري الميكروبي، تتغذى على الاوكزالات السامة وبذا تخلص الجسم منها. البكتريا سالبة لصبغة كرام، ولا تكون الابواغ ولاهوائية المعيشة، ويمكن عزلها من الغائط البشري، ولدراسة البكتريا من النواحي الجزيئية، تم جمع ع | Oxalobacter formigenes is one of the human microbiome, uses the toxic oxalate for their growth (Oxalotroph), and helps the body to get rid of excess oxalate. The bacterium is Gram negative, non - spore former, obligate anaerobe, and can be isolated from human stool.To carry out molecular studies on this bacterium, 50 samples of human stool were collected from subjects with wide range of ages (3 - 63 years) and different genders, subjects were healthy and have no medications and especially antibiotics for at least for 3 months. Genomic DNA was extracted with special kit, to amplify certain housekeeping genes, these are 16S rRNA (3' end) segment having the anti - Shine - Dalgarno sequence (ASD), and the oxc gene responsible for production of oxalyl - CoA decarboxylase (middle region responsible for active site at where the Thiamin pyrophosphate binds, the enzyme co - factor). It was possible to amplify the 16S rRNA gene in 46 sample out of 50 (92%), and 7 samples out of 50 (14%) for oxc gene. Analysis and alignments of 16S rRNA sequences put the Iraqi samples in the O. formigenes i.e., the same species depending on Distance Score of alignments, it has been found that ASD of bacteria is conserved and represented by the sequence " 5' CGCGGUGAACGUUCCCGGG3' " in the studied samples and in strains found in the International Databases. Segment of oxc (~ 600 bp) translated into proteins (190 - 194 amino acids) were aligned with oxc protein of O. formigenes (M77128) reference strain, it has been found that this segment similar to the TPP binding site and characterized by its high hydrophobicity
