000			02450nam a2200181Ia 4500
005			20151005151135.0
008			150525s2011 xx 000 0 und d
041			_aeng
082			_a1375,T
100			_aSaqib Hussain _97929
110			_cMr. Tanveer Hussain _96615
245			_aSequence Analysis Of Shiga Toxin 1 And Shiga Toxin 2 Genes Of Escherichia Coli O157: H7 Isikates From Lahore
260			_c2011
502			_aEscherichia coli is normal inhabitant of all the animals and human beings. The Sorbitol non fermenting E. coli strains were detected in milk, beef and fecal samples collected from different areas of Lahore. White colored colonies of each positive sample on Sorbitol MacConkey's Agar (SMA) contained gram negative, rods with round ends non spore former Escherichia coli. Each of the isolates was sorbitol non fermenter, lactose fermenter, indole positive, Methyl Red positive, Voges Prauskaur negative and citrate negative. Each of the isolate was further characterized using polymerase chain reaction (PCR) for the presence of shiga toxin 1 and shiga toxin 2 genes. Bacterial DNA was extracted easily by boiling method when the isolates were grown on Sorbitol MacConkey's agar at 37°C for 12-24 hours. The DNA was recovered when the culture was boiled for 5-10 minutes. The isolated DNA when amplified using Stxland Stx2 specific primers showed that 68.5 percent samples were positive for Stxl and 54.2 percent for Stx2. The stx 1 and stx2 PCR products were subjected to sequencing. The resulted sequences when aligned with the reference sequence through Basic local alignment tool it showed that the shiga toxin 1 and shiga toxin 2 gene sequences are conserved and showed high similarity in their nucleotide structure. Despite of having high similarity in their nucleotide structure some haplotypes were also obtained showing single nucleotide polymorphism. Phylogenetic analysis made among local isolates and also with reported sequences from all over the world by using bioinformatics software to see the genetic similarities and difference between them. The data produced showed some highly conserved sequences and SNPs as well that will be quite useful for further applications in diagnostics and biotechnology applications in future.
650			_aInstitute of Biochemistry & Biotechnology _95022
700			_aProf. Dr.Masroor Elahi Babar _96616
942			_cTH
999			_c3088 _d3088