Isolation Of Colonic Flora From Swine Institine And Expression Of Transgenic E.Coli In It

The 26 bacterial strains were isolated from jejunum and ileum of healthy and grown-up swine with some selective media .According to its shape of colony ,Gram stain and some biochemical characteristics as well as sugar fermentation reaction ,the preliminary identification was done, namely there are 5 strains of L.plantarum, 4 strains of L.fermentum, 1 strain of L.acidophilus and 6 strains of E.coli.The kind of dominant colonic Lactobacillus in jejunum and ileum was determined.So the researth provided a good foundations for expressing of transgenic E.coli(?-mannase) in swine effectively and reforming of the engineering bacteria and making probiotics and microecologics. The BL21-PT002-ManI/PUC18-Kil,YS- PT002-ManI/PUC18-Kil,BL21-PUC18-GFP and YS- PUC18-GFP have been established by transforming plasmids PT002-ManI,PUC18-Kil and PUC18-GFP(Green Flourescent Protein)to YS and BL21. The bacteria relatively accustomed to expression in vitro has been selected by researching characteristics of growth and expression on YS and BL21 with GFP characters and gene examination techniques. After swines have been selected to make cannulas at duodenum and ileum,the bacteria have been made microcapsule and then blent with others to become the pouring liquids to pour into swines through the cannula. The specimens have been examinated with colony PCR and ELISA .