Dissertation > Excellent graduate degree dissertation topics show

Construction of Genes Library and Analysis of Genes Ecoding Exported Proteins of Avian Pathogenic Escherichia Coli

Author: TanShuang
Tutor: YuXuPing
School: Zhejiang University
Course: Preventive Veterinary Medicine
Keywords: Escherichia coli gene encoding exported protein selective clone gene library
CLC: S852.61
Type: Master's thesis
Year: 2009
Downloads: 6
Quote: 0
Read: Download Dissertation

Abstract


Pathogenicity Escherichia coli infects a wide range of animal species and human.It has been reported that outer membrance protein especially those adopt regulatory expression mode is dominant immune antigen playing an important role in epidemic diseases, preventive.pMB-Ara-T vector was selected to clone genes encoding exported proteins of Escherichia coli 02 strain, pMB-Ara-T is a selection cloning vector containing the control promoter for transmembrane protein.It used to clone the special promoter, special conditions to be induced expression (in vitro culture conditions generally did not express) transmembrane protein gene. pMB-Ara-T is a vector in nature of T vector,can ligase directly with the PCR product.We use 30N8 and Escherichia coli 02 genomic DNA with a mixture of Kelnow extension as a template,OL30 as a primer,the PCR products amplified on the size of 200 bp～2000bp.Plasmid pMB-Ara-T digested by the XcmI and purified then ligase with the rPCR product and purified, transformed into competent DH5α,take 20ul from 1 ml of liquid conversion coating on the Kan resistant plate,grow 350 clones,afer ten transform get about 2×105library, the colony PCR analysis showed that the rate inserting more than 92 percent, fragment size between 250 bp and 500bp.The genmone were labled on Amp+Kan+l%arabinose and then on Amp+Kan plate.As a result,318 clones and 29clones were obtained. Sequencing,homology analysis that 318 clones were highly homology with 276 genes of E.coli 02 and the 29 clones were highly homology with 27 genes. Analysing all of the 318 clones, SignalP results demonstrated that 318 clones have the SP structure and the fragment was inserted as part of a ORF with the fusion of ampicillin genes; TMHMM results demonstrated that 78 clones have the more than one transmembrane structures; Analysing the 276 genes,144 genes carrying the complete promoter structure located in the upstream of starting codon.115genes were predicted to encode exported proteins with special needs induced by the secretion of gene expression, and the last 27 genes were predicted to encode exported proteins with regulatory expression mode.Gene Ontology results demonstrated that most of the genes were related to the transport, metabolic process, cell outer membrane and protein binding.Experimental results show that pMB-Ara-T can directly clone PCR products and screen the coded signal sequence of the transmembrane protein gene.The secretion protein and membrane protein genes by Screening provide a favourable basis for further study of toxicity, immunogenicity and other related transmembrane protein and gene expression.

Related Dissertations

  1. Cloning and Expression of Growth Hormone Gene in Cobia (Rachycentron Canadus),Q786
  2. Expression of VP2 Gene of Infectious Bursal Disease Virus and Preparation of Monoclonal Antibodies Against the Recombinant VP2 Protein,S852.65
  3. Isolation and Identification of Avian Pathogenic Escherichia Coli from Chicken and Characterizations of Its Virulence-Associated Genes,S852.61
  4. Cloning and Expression of Organic Solvent Tolerant Lipase Gene from Staphylococcus Saprophyticus M36,Q78
  5. Study on Mixed Strain for Liquid State Fermentation of Animal Blood,S816.4
  6. Antimicrobial Resistance Surveillance of Escherichia Coli in Jiangsu; and Discussion on the Resistance Monitoring and Surveillance Program,S854
  7. Inactivation of Escherichia Coli in Fresh Cucumber Juice and a Shelf Life Study Using High Hydrostatic Pressure,TS255.5
  8. Identifying and Clonging the cry30Ga2 Gene from B. Thuringiensis Isolate S2160-1,Q78
  9. The Research on Drug-resistance, Genotype and Related Diseases of Extended Spectrum-lactamases Escherichia Coli,R446.5
  10. The Pollution of Escherichia Coli O157: H7 and the Risk Assessement about Cold Dishes,R155.5
  11. Study on Characteristics of Resistance in Piglet-associated Escherichia Coli Against Beta-lactam Antibiotics in Chongqing,S852.61
  12. The Study of Resistance to Fluoroquinolones in Escherichia Coli,R96
  13. Study of CTX-M-14 Extended Spectrum β-lactamases in Producing ESBLs Escherichia Coli,R450
  14. Research on the Resistance of Intestinal Flora in Mice and Klebsiella Pneumoniae to Fluoroquinolones,R446.5
  15. The Expression and Purification of Peptide Deformlyase and the Study of Its Inhibitors,R392
  16. A Study on the Repetitive Extragenic Palindromic PCR Fingerprint Analysis As A Microbial Source Tracking,R123
  17. Studies on the Secretory Expression of Arginine Deiminase,Q78
  18. Detection and Genotype Analysis of Extended-spectrum β-lactemases(ESBLs)Producing Escherichia Coli Isolated from Chenzhou Region,R440
  19. Surveillance of Antimicrobial Resistance and Detection of Integron Gene Cassettes among Escherichia Coli Isolates from the Intestinal of Animals,S852.61
  20. Construction of Recombinant Catalase Engineering Strain and Its Fermentation Optimization,TQ925
  21. Establishment of Multiplex PCR for Staphylococcu Saureus, Streptococcus, Salmonella Spp and Escherichia Coli,S855.1

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Pathogenic bacteria
© 2012 www.DissertationTopic.Net  Mobile