Dissertation > Excellent graduate degree dissertation topics show

Cloning and Expression of Main Structure Protein Genes of Taura Syndrome Virus in P. Vannamei Shrimp

Author: LiuTang
Tutor: ChenLiang
School: Xiamen University
Course: Cell Biology
Keywords: Taura virus Clone Litopenaeus vannamei
CLC: S945.4
Type: Master's thesis
Year: 2008
Downloads: 89
Quote: 0
Read: Download Dissertation

Abstract


Litopenaeus vannamei is the world's most important economic farmed shrimp varieties worldwide, a large number of breeding. Their disease worsening, shrimp viral disease outbreaks caused huge economic losses to the world shrimp farming industry poses a great threat to the sustainable development of marine resources, has become the world shrimp disease research hotspot in the field of shrimp viruses a. Taura syndrome virus disease affect Litopenaeus vannamei production is the most important one, the fatality rate as high as 90% or more. Taura syndrome was initially found only in Ecuador, South America, by transporting infected with the virus of the larval shrimp and broodstock spread of the virus to the Americas, and further spread to the whole world. Taura virus diagnostic method organized pathology cDNA probe in situ hybridization, RT-PCR method, but these methods do not meet the rapid quarantine requirements of the port sector, an urgent need for a quarantine of a fast and accurate method . In this study, according to the the GenBank, peach Pull virus whole genome sequence design amplified the virus major structural protein genes the vp1, vp2, the Major, vp3 the corresponding primer material to the onset of Litopenaeus vannamei extracted total RNA as template, using RT- PCR amplified target gene, respectively, to obtain the molecular weight 1164bp, 849bp and 507bp gene fragment, fragment size is consistent with expectations. Connection will purpose fragment pTA2 carrier, introduced into E. coli DH5α positive recombinants pTA-VP1 the PTA-VP2, PTA-vp3. Blast analysis of the target fragment sequencing results show that has been successfully related protein gene fragment. This positive recombinants and pET-28a vectors were digested recombinant expression vector pET-VP1 pET-VP2 and pET-VP3, recombinant after restriction analysis and sequencing introduced into E. coli BL21 (DE3), IPTG was induced expression by SDS-PAGE detection. SDS-PAGE electrophoresis results showed that at 37 ℃ 0.5mmol/dm 3 of IPTG induces the expression of recombinant protein expressed in the form of inclusion body recombinant proteins VP1, VP2, VP3 the size were 48 kDa, 36 kDa and 24kDa . Refolded recombinant protein was purified using nickel column, the purified recombinant protein was used to immunize mice to produce polyclonal antibody and Western immunoblot reaction results showed that the antibodies obtained react specifically with recombinant protein Dot-ELISA results show that the obtained antibody react specifically with shrimp disease lesions. The results of this study for the development of the Taura virus rapid test kit to lay the foundation.

Related Dissertations

  1. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  2. Cultivation and Effects of Environmental Factors on Carrying WSSV Virus of Litopenaeus Vannamei,S945
  3. Comparison on Immunoprotective Effect of Bacteriophages of Vibrio Alginolyticus and Vaccines of Bacterial Ghost,S945
  4. Cloning and Characterization of BAFF Gene from Whitespotted Catshark and Crucian Crap,S917.4
  5. GIH cDNA Cloning and Mrna Expression in Litopenaeus Vannamei,S917.4
  6. Cloning Andanalysis of NAC Transcription Factor Gene in Tobacco(Nicotiana Tabacum L.),Q943.2
  7. Oshima Ye Lu Ju CcSOS1 Cloning and Expression Analysis,S682.11
  8. The Transmembrane Fusion Protein of MnSOD Clone Expression and Function Study,Q78
  9. Günther Myostatin gene pathway gene cloning and expression,S917.4
  10. Functional Analysis of Plant Asparagine Synthetase Gene in Regulation of Disease Resistance,S511
  11. Determination of Digestibility of Selected Protein Sources and Evaluation of Feed Formulation Based on Their Availability for Pacific White Shrimp, Litopenaeus Vannamei,S963
  12. Study on Variation in Phenotypic Characteristics and Photosynthetic Physiology of Young Camellia Oleifera Clones of Group Gan,S794.4
  13. Cloning and Expression Analysis of Hsp70 Gene from Freshwater Planarian Dugesia Japonica,Q78
  14. Sequence Analyzing and Prokaryotic Expression of ORF2 Gene in the Strain of Porcine Circovirus Type 2 Isolated from Lanzhou,S852.65
  15. Cloning Gonald-inhibiting Hormone Gene in Eriocheir Sinensis,S917.4
  16. Cloning and Expression Analysis of Vitellogenin Gene from Actias Selene Hubner,S885.9
  17. Researching on Transferring Human Insulin Gene into Chicken,S831
  18. Phenylalanine Ammonia-lyase Gene from Camellia Oleifera Full-length Gene Clone and Expression in Prokaryotic and Eukaryotic,S794.4
  19. The Cloning and Expression of INHIBIN, Preparation and Identification of McAbs,R392.1
  20. Construction of Gene Engineered Bacteria of Xylanase and Its Recombined Enzymatic Properties,Q78
  21. Study on the Nitrogen and Phosphorus Budget and Production Performances in Intensive Pond Culture of Shrimp,S968.22

CLC: > Agricultural Sciences > Aquaculture, fisheries > Fisheries Protection > Pest and Disease Control of the crustaceans > Pest and Disease Control of a variety of shrimp
© 2012 www.DissertationTopic.Net  Mobile