Dissertation > Excellent graduate degree dissertation topics show

Construction of Yeast Two-Hybrid cDNA Library from 18day Schistosoma Japonicum and Screen the Factors of Interactive with SjGCP

Author: LiMian
Tutor: ZhangLongXian；JinYaMei
School: Henan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Schistosoma japonicum Yeast two-hybrid cDNA library Gynecophoral canal protein Filter
CLC: S852.735
Type: Master's thesis
Year: 2009
Downloads: 63
Quote: 0
Read: Download Dissertation

Abstract


Japan schistosomiasis (Schistosomiasis japonica) is a serious zoonotic parasites of a hazard. Schistosomiasis endemic in our country is mainly caused by schistosomiasis japonica in Chinese mainland strain. Mature females produce a large number of eggs is the main reason for a lot of pathological changes caused by schistosomiasis japonica discharge eggs is also the source of infection disease occurred. If the separation will be paired females and males, the females will stop spawning and degraded to the status of one kind is not fully developed, if re paired, it is still able to continue to mature. Hold the the female canal protein (SjGCP: Schistosoma japonicum gynaecophoral canal protein) present in the adult surface the male surface and confined to the site of action in the pairing directly - expressed gynecophoral canal. Interestingly, the transcription gynecophoral canal protein expression peaked in the male and female worms pair while. This indicates that the, gynecophoral canal protein plays a potential role in the interaction and stimulation of the female of the male and female worms mature. Therefore, in-depth study of protein gynecophoral canal to expose the schistosome mature mechanism is particularly important. For this purpose, we have constructed a cDNA library of 18-day-old the Schistosoma japonicum yeast two-hybrid and initial screening with gynecophoral canal protein. 18-day-old of Schistosoma japonicum yeast two-hybrid cDNA library constructed: select the 18-day-old worms of Schistosoma japonicum parasite total RNA was extracted using Trizol method Oligo (dT) primer to one strand cDNA was synthesized by reverse transcription, and then to 5 ' amplimer and. 3'amplimer as a primer, the LD-PCR amplification, so as to obtain double-stranded cDNA with CHROMA SPINTE-400, column purified double-stranded cDNA hybridized with yeast (Yeast mating) method with pGADT7-Rec transformed to feelings state yeast cells AH109, ??both homologous to the yeast cells composed of annular replication activity of cDNA library plasmid collected missing leucine (SD /-Leu) culture plate all clones, namely the 18-day-old Schistosoma japonicum yeast two hybrid cDNA Library bacteria. The transformant cell density is greater than 2 × 107cells/mL, storage capacity of 2.5 × 106 pfu / mL of the length of the double-stranded cDNA fragment inserted between 0.25 ~ 2.0kb library recombination ratio was 99%, the library has a good representative. Six pairs of specific primers can fish from the library to the adult Schistosoma japonicum genes. The test using the yeast two-hybrid technology was successfully constructed a cDNA library of Schistosoma japonicum 18-day-old adult. Used in the yeast two-hybrid system gynecophoral canal protein fusion plasmid and validation: hold the encoded protein gene of estrogen ditch complete open reading frame as the target protein is cloned into the pGBKT7 plasmid to construct recombinant vector pGBKT7-SjGCP the pGBKT7 -SjGCP Go yeast Y187 strain competent cells. Since the transcriptional activity detection, protein expression and toxicity testing results show that the the protein applies SjGCP target in the yeast two-hybrid system. Screening factor gynecophoral canal protein interactions: yeast hybridization (Yeast Mating) Schistosoma japonicum 18-day-old yeast two-hybrid cDNA library screening, the director of the positive clones (locus coeruleus) repeated passage screening, we obtained 330 clones. Randomly selected five clones extracting yeast plasmid, and electricity conversion method to go to E. coli DH5a strains were sequenced. We got five Schistosoma japonicum EST sequences,: SJCHGC0566829, SJCHGC09129, SJCHGC 00694 SJCHGC0607 and SJCHGC05265, respectively twinfilin protein gene expression Schistosoma japonicum, collagen protein gene, HSP60 protein genes and unknown genes. 3 has got the sequence of the five clones were selected randomly cloned using the method of co-transformation and analysis of its interaction in yeast.

Related Dissertations

  1. Satellite Attitude Determination Based on Gyro and Star Sensor,V448.2
  2. Design and Processing of Hybrid Modulated HF Radar Waveform,TN958.93
  3. Screening Molecular Targets of Phytophthora Sojae RXLR Effectors,S435.651
  4. Analysis of Infrared Spectral Features and Identification of Al-responsive Genes in Small Black Soybean with Aluminum Stress,S529
  5. Active Power Filter and Its Application in Distribution Network,TN713.8
  6. Yeast Two-Hybrid Screening of 14-3-3-Interacted Proteins During Early Cotton Fiber Development,S562
  7. Molecular Cloning, Prokaryotic Expression and Characterization of a Novel Trypsin Inhibitor in the Frog, Lepidobatrachus Laevis,Q78
  8. Construction and Primary Analysis of Differentially Expressed cDNA Library of Alfalfa by Using Suppression Subtractive Hybridization under the High Temperature,S541.9
  9. Purification of Recombinant Schistosoma Japonicum Protein SjGST and Preparation and Characterization of the Monoclonal Antibody,R392
  10. Cloning and Preliminary Functional Analysis of GmCASc Gene in Soybean (Glycine Max(L.) Merrill),S565.1
  11. Treatment of Effluent Water from Bio-flocculation Adsorption Technology by Biological Aerated Filter (BAF),X703
  12. The Value Simulation of the Filtration Process by Fiber Filter Materials,TB34
  13. A Novel Controller Design for Active Power Filter with Low Switching Loss,TN713.8
  14. Study on Pharmacokinetics and Spatial and Temporal Expression in Mice of DNA Vaccine against Schistosoma Japonicum,S855.91
  15. The Research and Design of Phase Conjugation Mixer in Retro-directive Array,TN773
  16. Degradation of Dibenzothiophene by Some Newly Isolated Bacterial Strains,TE621
  17. Experimental Study of the Performances of Air Filter Materials for Fine Particles Removal Utilized in Residential Buildings,TU834
  18. Research of Active Power Filter in Three-phase Four-wire System,TN713.8
  19. Research on the Application of Fundamental Magnetic Flux Compensation-type Series Hybrid Harmonic Suppression Device,TN713.8
  20. The Research and Design of Signal Processing Technology of the Optical Fiber Displacement Sensor Based on Intensity Modulation,TN911.7
  21. AUV Integrated Navigation Algorithm Study and System Implementation,U666.1

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Livestock parasitology > Veterinary worms school > Fluke
© 2012 www.DissertationTopic.Net  Mobile