Construction of feed additive probiotics for poultry by using genetic engineering techniques

ABSTRACT MSc THESIS CONSTRUCTION OF FEED ADDITIVE PROBIOTICS FOR POULTRY BY USING GENETIC ENGINEERING TECHNIQUES MELTEM AŞAN DEPARTMENT OF ANIMAL SCIENCE INSTITUTE OF NATUREL AND APPLIED SCIENCES UNIVERSITY OF ÇUKUROVA Supervisor: Doç. Dr. NUMAN ÖZCAN Year: 2002, Pages: 77 Jüri: Doç. Dr. Numan ÖZCAN Prof. Dr. Hasan Rüştü KUTLU Yrd. Doç. Dr. Mehmet Sait EKİNCİ An integration vector (pLAlL) of Lactobacillus acidophilus A-161 carrying mixed linkage p(l,3-l,4)-glucanase were developed. 3 kbp EcoRI-EcoRI random fragment of Lactobacillus acidophilus A-161 genome were cloned in CMoramphenicol resistant gene oî m Escherichia coli replicative vector of pBR325 to construct pLAl integrative plasmid. In addition, 1,8 kbp Hindm-BamHI fragment of a rumen bacterium Streptococcus bovis JB1 encoding |3(l,3-l,4>glucanase enzyme, located in pLIHc vector, were transferred into Tetracycline resistance gene of pLAl to create pLAlL. Probably, because of unoptimized electroporation conditions, it could not integrated into Lactobacillus acidophilus A-161 choromosome by electrotransformation. TL1R construct, consisted of Escherichia coli I Streptococcus shuttle vector pTRWIO and mixed linkage 0(l,3-l,4)-glucanase gene, was succesfully electrotransformed and expressed into Streptococcus thermophilus FI8976 and in Lactococcus lactis IL1403 strains. SDS-Lichenan-PAGE jels showed no sing of protease degredation, in both strains of 26 kDa enzyme protein. Key Words: Lactic Acid Bacteria, Probiotics, Beta-glucanase, Subcloning rf<*