Dissertation > Excellent graduate degree dissertation topics show

Cloning and Overexpression of Goat Toll-like Receptor 2 Gene

Author: FengGuoXing
Tutor: PanQiuZhen
School: Heilongjiang Bayi Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Goat TLR2 Clone Gene Expression Fusion protein Overexpression of
CLC: S827
Type: Master's thesis
Year: 2011
Downloads: 55
Quote: 0
Read: Download Dissertation

Abstract


Toll-like receptors (Toll-like receptors, TLRs) are pathogen associated molecular pattern recognition receptors to activate innate immunity and acquired immunity through the identification of pathogenic microorganisms . Toll-like receptor 2 (TLR2) Toll-like receptor family to identify the type of ligand up to a receptor . Play an important role in many infections caused by pathogenic microorganisms , and therefore the the TLR2 research receiving more attention . The domestic sheep TLR2 research has not been reported . This study from the goats spleen tissue extraction of total RNA by RT-PCR technology , the TLR2 gene was cloned into the vector pGEM-Teasy . After sequencing analysis , cloned goat TLR2 connection to the pEGFP-N1 vector , building green fluorescent protein fusion vector pEGFP - of TLR2 -N1 . Subsequent fusion vector transfected 293T cells , the expression of the fusion protein was observed under a fluorescence microscope . Re- construct the recombinant vectors of the 3S-Loxp-TLR2 transfected Goat fibroblasts, the positive cells was purified using G418 selection . PCR detection of DNA extracted positive cells . Using real-time quantitative PCR method to detect the expression of TLR2 . The first cloned goat TLR2 gene , submitted to the GenBank accession serial number : GU984768.1 . By sequencing analysis , ORF is the gene 2355bp , encoding 784 amino acids , goat the TLR2 sequencing results will be obtained with other species have been published TLR2 sequence comparative analysis results show that the results of sequencing with sheep , cattle , the great Indian blue antelope similarity , pigs, horses, dogs, etc. , followed by chicken lower similarity . That in line with the characteristics of the TLRs : extracellular domain, a transmembrane region and intracellular region of three parts constitute cloned goat of TLR2 protein structure prediction . PEGFP-TLR2-N1 fusion vector constructed in this study , 293T cells were transfected using the calcium phosphate method , the expression of green fluorescent fusion protein can be targeted to the plasma membrane . Fibroblasts transfected with the recombinant vector of the 3S-Loxp-TLR2 goat by G418 PCR identification of green fluorescent protein fluorescence microscopy , and real-time quantitative detection TLR2 expression means of identification , goats , TLR2 gene in goat fibroblast cells the excessive expression, and the cells were in good condition . Lay the foundation for further study the function of the goats TLR2 mechanism as well as the production of somatic cell cloned transgenic goats .

Related Dissertations

  1. Research on Combinatorial Regulation of Multiple Transcription Factors,Q78
  2. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  3. The Expression Survey of BMP Signalling Pathway in the Human Embryonic Tooth Germ,R78
  4. Cloning and Characterization of BAFF Gene from Whitespotted Catshark and Crucian Crap,S917.4
  5. Study on Synthesis and Physiological Function of N-carbamoyl-glutamate,R914
  6. Functional Analysis of Two Rice Nitrate Transporter Genes OsNRT1.1a and OsNRT1.1b,S511
  7. Coparative Analyses of Carbon and Nitrogen Metabolism of Flue-cured Tobacco Growing in Henan and Yunnan,S572
  8. Research on Influnce of Silicon and Sulfur on Arsenic Uptake and Accumulation of Rice (Oryza Sativa L.),S511
  9. Study of Chicken Δ~6 Fatty Acid Desaturase Gene’s Promotor Region Polymorphism and Gene Expression,S831
  10. Cloning Andanalysis of NAC Transcription Factor Gene in Tobacco(Nicotiana Tabacum L.),Q943.2
  11. The Expression of CPV-2 VP2 Gene in SF9 and Development of Indirect Elisa for Serum Antibodys,S852.65
  12. Cloning and Molecular Analysis of Lovastatin Biosynthesis Related Genes from Monascus Purpureus,TQ927
  13. Comparison of Gene Expression Data Cluster Methods and Gene Network Construction for Phytophthora Sojae Genes,S435.651
  14. Functional Identification of Two Tomoto Phosphater Transporter Genes LePT1 and LePT2 in Rice,S511
  15. Analysis for Intron 1 and 2 of Rice Transformation to Promoter Activity,S511
  16. Research on Characteristics of Growth, Development and Material Metabolism of Flue Cured Tobacco Leaves in Huili, Sichuan,S572
  17. Studies on Extracting Mitochondrial DNA and Gene Expressions in Brassica Napus L,S565.4
  18. Functional Characterization of Wheat Na~+ /H~+ Antiporter TaNHX2 and Study of TaNHX2 C- Terminus Domain,S512.1
  19. Evaluation on Forage Quality and Biomass Energy Characters of Inbred Vegetative Lines of Napier Grass,S543.9
  20. Identification and Analysis of miRNA and Flower Development Specific Genes in Wheat,S512.1
  21. Molecular Characterization, Expression, and Promoter Methylation Status of the Bovine Synaptonemal Complex Protein 3 Gene,S823

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Goat
© 2012 www.DissertationTopic.Net  Mobile