Dissertation > Excellent graduate degree dissertation topics show

Construction and Identification of Lentivial RNA Interference Vector of Sheep myostatin Receptor Gene

Author: ShengPengCheng
Tutor: ZhuRuiLiang
School: Shandong Agricultural University
Course: Preventive Veterinary Medicine
Keywords: RNAi shRNA myostatin lentiviral qPCR
CLC: S858.26
Type: Master's thesis
Year: 2011
Downloads: 31
Quote: 0
Read: Download Dissertation

Abstract


Objective: To construct the lentiviral RNA interference (RNAi) vector of sheep myostatin gene and evaluate the effects of silencing myostatin gene expression by shRNA. Methods: Firstly retrieve purpose gene sequences in the Genebank to get myostatin sequence of sheep. Design and synthesize four pairs of oligonucleotide sequence target myostatin, and synthesize high-expression vector according to the ambion shRNA design system. Synthetic whole gene sequences, which is put into pMD-18T carrier. Find mutation points by sequencing, repair the mutating gene sequence through the overlapping PCR, then use XhoI and EcoRI enzyme cut the sequent, and connect to pCDNA3.1(+).high-expression vector is constructed by sequencing. Transfect shRNA and high-expression vector into HEK293cells, the gene silencing efficacy of the four targets was compared by real-time PCR, ShRNA - 86C’s interference efficiency is best, gene silencing efficiency is 60.13%, ShRNA - 86C is screened and vector pDONR221 execute BP recombination reaction, so to obtain an entry vector containing interference sequence. Then the entry vector containing interference sequence and the pLenti6/BLOCK-iT-DEST vector execute LR recombination reaction to get the lentiviral expression vector with interference sequence. qPCR detection of the physical titer of the virus. Results: Successfully constructed Lentivector-mediated-shRNA, and the titer is 8.8×10~9TU/ml. Provide a stable infected cells vector to study the impact of myostatin gene on muscle growth.

Related Dissertations

  1. Improvement of the Resistance of Nicotiana Tabaccum to Virus by RNAi and Initial Establishment of Genetic Transformation System of L.regale,S435.72
  2. Study on Roles of Diacylglycerol Kinase Gene Family in Rice under Xylanase and NaCl Treatments by Transient RNA Interference System,S511
  3. Study on the Generation of Transgenic Embryos of Cattle and Mouse with Lentiviral Vector,S814.8
  4. Expressive Characterization of Transcription Factors in Cotton Fibre Initiation and Cloning and Characterization of Three Transcription Factor Genes in Gossypium Hirsutum L.,Q943
  5. Toxicities of Several Newly-Typed Insecticides to Colorado Potato Beetle, Leptinotarsa Decemlineata (Say) and Molecular Cloning of Two Kinds of Targets,S435.32
  6. The Genes Expression Analysis of Cotton Fiber Initiation Stage and Characterization of Three New Genes in Gossypium,S562
  7. Construction of Blg Knockdown Cell Lines by Using Lentiviral Vector-Mediated Ran Interference,S827
  8. Expression of β-Catenin in Pig’s Ovary and the Effect of β-Catenin on Porcine Granulosa Cells Apotosis and Steroidogenesis Related Enzyme,S828
  9. Inhibition of Myostatin (MSTN) Expresstion in Primary Porcine Fetal Fibroblasts by Lentivector-mediated RNAi,S828
  10. Study on Genes Expression and Function of UDP-glucosyltransferases in the Silkworm, Bombyx Mori,S881.2
  11. Apoptosis Mechanisms Research in Interfering TIGAR Gene and Epirubicin Treatmnet in Hepatic Carcinoma Cells,R735.7
  12. Analysis of Antenna Protein of Silk Moth by Two -dimensional Electrophoresis,Q966
  13. An Experimental Study of Inhibition the Expression of Ppif Gene Reduce Renal Ischemia-reperfusion Injury in Rats,R363
  14. Effect of STAT1-shRNA-mediated Gene Silencing on Biological Characteristics in the Cervical Cancer Cell Line Hela,R737.33
  15. Günther Myostatin gene pathway gene cloning and expression,S917.4
  16. Functional Analysis of Plant Asparagine Synthetase Gene in Regulation of Disease Resistance,S511
  17. RNAi-mediated inhibition of MCF-7 cells expressing the VEGF gene,R363
  18. Cloning and Expression of Gallerimycin from Spodoptera Litura,S433
  19. Functional Analysis of Genes MaFKS and MaChsⅦ Related to Cell Wall Synthesis in Metarhizium Acridum,S476.1
  20. Selective Detection of Viable Pathogens in Water Using Propidium Monoazide Combined with qPCR and Its Application,R123.1
  21. The Reseach of Controlling Aphis Gossipy by Transgenic Technology,S188

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Sheep
© 2012 www.DissertationTopic.Net  Mobile