Exploration Of Bacterial Aquaporin And Its Physiologic Function

Part I Explore bacterial aquaporin in 11 pathogenic bacteriumObjective: find bacterial aquaporin in S.aureus,S.epidermidis,Ssa-rophyticus,E.coli,S.dysenteriae,S.paratyphib,H.pylori,P.aeruginosa,M.chel-onei,P.vulgaris,C.albicans with the degeneracy PCR. The finding will make the basic foundation for its physiologic function.Methods: Chromosomal DNA from 11 pathogenic bacterium w-as used as template in a PCR reaction with degenerated primers designed from the two conserved'NPA'sequences of the MIP family. We found 400bp sequence in two pathogenic bacterium, then BLAST on web.Results: we found Glp-F in E.coli the same as abroad, we new discover Glp-F in S.dysenteriae and no report at present. Conclusions: we used Chromosomal DNA from 11 pathogenic bacterium as template in a PCR reaction with degenerated primers designed from the two conserved'NPA'sequences of the MIP family, and cone two same sequence as Glp-F, it give a foundation for research its physiologic. Part II The aquaporin gene Glp-F of S.dysenteriae is induced in hyper osmotic conditionsObjective: S.dysenteriae growth in Nutrient Agar which have different osmotic conditions, determinate OD600 at 24,48,72,96 respectively. The reverse transcription assay was used to quantify the levels of Glp-F mRNA in different osmolarity conditions.Methods: Basic media made as Nutrient Agar. Media-N were made hypertonic by adding NaCl to increase the concentrations up to 125, 250, 500 or 750 mM. Media-N were made hypotonic by diluting them with water (50 % and 33 %). Media-H were made hypertonic by adding NaCl and HgCl2 to increase the concentrations up to 125, 250, 500 or 750 mM. Media-H were made hypotonic by diluting them with water (50 % and 33 %) and add 2mg HgCl2/80ml.Result: S.dysenteriae growth indifferent in media-n/h witch were made hypotonic by diluting them with water (50 % and 33 %) and in basic media. We found S.dysenteriae growth rapider in hypertonic media-N than growth in hypertonic media-H. The reverse transcription reveal that expression of more Glp-F m-RNA in hypertonic media-H than growth in hypertonic media-N.Conclusions: Glp-F of S.dysenteriae made significance role in hypertonic environment than in hypotonic media. Glp-F made the use of control osmotic stabilization in vivo under hypertonic media. Thes e results indicated that the expression of Glp-F was regulated during the growth curve and induced in hyperosmolar conditions.