الفهرس 
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Abstract
ABSTRACT
The present investigation aims to study the impact of some genetic treatments (using mutation by EMS and protoplast fusion) on the probiotic activities of Saccharomyces boulardii yeast strain. As well as S. boulardii whole cells were transformed using yeast recombinant plasmid (pRSH-Ins) and enzymatic amplification of genetic material using PCR technique. Most experiments of this investigation were conducted in microbial genetics laboratory National Research Center. While some experiments were carried out in the microbial genetic laboratory, Genetics Department, Faculty of Agriculture, Cairo University, Giza, Egypt. The present study was conducted during 2004- 2009.
Saccharomyces boulardii was used to examine the changes in different points; the ability to tolrate bile salts, bioaccumulation of zinc, selenium, antimicrobial activity and the productivity of vitamine B6, and some important character such as insulin production ; following EMS as chemical mutagenic agent, protoplast fusion and transformation as a genetic engineering method . Induction of mutation in S. boulardii was carried out by (1.17 g/ml) to take 0.250, 0.500, and 1000 µl ml-1 results revealed that the survival percentages was decreased by increasing the doses of the amount of EMS where as the survival percentage was 7.9% at exposure dose 1000 µl /ml-1. eight mutants were isolated as auxotrophes and their nutritional requirements were determined. S. boulardii and their resulted mutants were tested for 5% bile salts tolerance, and selection of high zinc yeast The results showed that mutants S.b M7 tolerated 5% bile salt and mutants S.b M6 is a highly zinc yeast. S.b M8 highly selenium bioaccmulation, S.b M7 is a highly antimicrobial activity biotherapeutic yeast S.boulardii strains by interspecific protoplasts fusion(3.54%) . Protein banding patterns SDS PAGE from w.t and their mutant show specific band of the different EMS treatment and fusant may be referring to the occurred chane of different character of the S.boulardii. Genetic construction of new strains using yeast recombinant plasmid (pRSH-Ins) and enzymatic amplification of genetic material using PCR technique was used in the present work to detect the human pro-insulin gene corresponding sequence in the newly S. boulardii cells. Four successive amplifications were obtained using the primer set (P1). When the amplified products were run on 1 % agarose gel electrophoresis at ~264bp.
Key words: Mutation, EMS, S.boulardii, protoplast fusion, protein fingerprinting, Bile salt, Zinc, Selenium, Antimicrobial activity,Vitamin B6, Transformation, Proinsulin gene