TAILIEUCHUNG - Báo cáo khoa học: "Expression of apxIA of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Expression of apxIA of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae | J. Vet. Sci. 2003 4 3 225-228 JOURNAL OF Veterinary Science Expression of apxIA of Actinobacilluspleuropneumoniae in Saccharomyces cerevisiae Sung Jae Shin Jong Lye Bae1 Young Wook Cho Moon Sik Yang1 Dae Hyuk Kim1 Yong Suk Jang1 and Han Sang Yoo Department of Infectious Diseases College of Veterinary Medicine and School of Agricultural Biotechnology Seoul National University Seoul 151-742 Korea 1 Division of Biological Sciences and the Institute for Molecular Biology and Genetics Chonbuk National University Chonju 561-756 Korea Actinobacillus pleuropneumoniae is an important primary pathogen in pigs in which it causes a highly contagious pleuropneumoniae. In our previous study apxIA gene amplified from A. pleuropneumoniae Korean isolate by PCR with primer designed based on the N- and C-terminal of the toxin was cloned in TA cloning vector and sequenced. The nucleotide sequences of apxIA gene was reported to GeneBank with the accession numbers of AF363361. Identity of the Apx IA from the cloned gene in E. coli was proved by SDS-PAGE and Western blot. Yeast has been demonstrated to be an excellent host for the expression of recombinant proteins with uses in diagnostics therapeutics and vaccine productions. Therefore to use the yeast as a delivery system in new oral subunit vaccine apxIA gene was subcloned into Saccharomyces cerevisiae and identified the expression of Apx IA protein. First apxIA gene was amplified by PCR with the primers containing BamHI and SalI site at each end. Second the DNA digested with BamHI and SalI was ligated into YEpGPD-TER vector and transformed into S. cerevisiae 2805. Third after identification of the correctly oriented clone the 120-kDa of Apx IA protein expressed in S. cerevisiae 2805 was identified by SDS-PAGE and Western blot. Key words Actinobacillus pleuropneumoniae apxIA expression Saccharomyces cerevisiae Introduction Porcine pleuropneumoniae caused by Actinobacillus pleuropneumoniae is a contagious fibrinous hemorrhagic .

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