上海地区猪链球菌2型的流行病学调查和其PGK基因缺失株的构建.pdfVIP

Epidemiological Survey of SS2 in the Shanghai Area and the Construction of the PGK Gene Deletion Strains Abstract Streptococcus suis disease is caused by Streptococcus suis.It is an important zoonotic disease,especially streptococcus suis type 2,not only the most pathogenic in pigs and specific populations and death,to human pubic health and pig industy have a tremendous impact. But so far, the pathogenesis is not very clear. Therefore, this study on the basis of the epidemiological investigation and analysis of drug resistance, to be through gene knock addition to the technical building gene deletion strains, and to lay the foundation for further study the pathogenic mechanism of the Streptococcus suis. In this paper carried out the following studies. Suspected streptococcus suis type 2-infected pigs of whole blood,internal organs and faeces collection of clinical symptoms, the samples for bacteriologicalisolated and purified,the use of the identification of double PCR method,according to test results,select all virulence factors-positive strains,application of SPF Bama miniature pig as an animal model for pathogenicity experiments,filter out the virulent strain,available for the following experiments.Finally,paper diffusion strain of Streptococcus suis type 2 clinical isolates to determine its resistance. In this study,homologous recombination were selected,on both sides about the upstream and downstream of the PGK gene of 500bp and 700bp sequence as homologous arms and the screening of Cm resistance gene were cloned into plasmid vector PUC18.Be connected to by the EcoR I/ Sph I digestion pUCl8-PsCmPa,recovery insert tablets degree,will be recycled form the fragment was cloned into the EcoR I/ Sph I double digested shuttle vector pSET4s ligation products transformed into E.coli DH5a extract the plasmid was transformed into the SH08 competent cells,the recovery after 28℃ and 37℃ plasmid loss.the use of PCR and resistance were scr