Dissertation > Excellent graduate degree dissertation topics show

Molecular Cloning and Characterization of Porcine Intelectin2 Gene

Author: ZhangJuan
Tutor: YangZaiQing
School: Huazhong Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Meishan pigs intelectin2 characterization regulation of gene expression dual-luciferase reporter assay system SREBP-1c
CLC: S828
Type: Master's thesis
Year: 2010
Downloads: 12
Quote: 0
Read: Download Dissertation

Abstract


Meishan breed is famous for its fecundity, it is an important breed resource all over the world. There are still many puzzles on the molecular mechanisms of fat deposition and ectopic fat deposition, lipid metabolism to solve. So functional research of important genes in this breed would not only be necessary for the further genetic improvement in accumulating and exploring the gene resources and breeding, but also for clarifying the molecular mechanisms of fat deposition and ectopic fat deposition, lipid metabolism and so on.Intelectin was first reported as a newly discoved gene which specially expresses in visceral adipose tissue, it cause widespread attention as an adipokine secreted by omental adipose tissue. In humanity, it is the link between obesity, insulin resistance, cardiovascular disease and metabolic syndrome. There are two isoforms:intelectin 1、2, both of the two gene expression were decreased with obesity and were highly correlated with each other in visceral adipose tissue. Besides, as intelectin has a fibrinogen domain, most researches focus on their numerous functions as lectin in intestinal, however, not many more researches on its molecular cloning and characterization in pig.In this study, we first reported here the intelectin2 CDs and 4 introns cloning and tissue distribution by qRT-PCR.We also had successfully cloned and analysised the 5’ flanking region of intelectin2, and further constructed to pGL3 luciferase reporter vector. By using dual luciferase assay system, the activity of the region was analysised in porcine IBRS-2 cells. To understand the expression and regulation mechanism of porcine intelectin2,the promoter pGL3 luciferase reporter vector had been transfected into mouse NIH3T3 with eukaryotic transcription factor pcDNA3.1-C/EBPa (CREB1, KLF15, PPARγ2, SREBP-lc). Dual luciferase assay system was used to detect the activity changes of promoter after transfected transcription factors. Our findings provide the basis for clarifying the regulation method and functions of porcine intelectin2. The main results are as follows:1. We had successfully cloned the intelectin2 CDs(1213bp) and 4 introns (ranging from 234bp to 1047bp).According to the CDs and several introns, we analysis the genomic structure of intelectin2 which consists of 7 introns and 8 exons, all of the exon-intron boundaries are agreed with the GT/AG rule. Porcine intelectin2 share 81%, 84% and 84% identity with intelectin2 gene of human, cattle and mouse, respectively. Tissue distribution by qRT-PCR indicated it is abundant in gut, with low expression in abdominal fat, and undetectable in several tissues.2. Porcine intelectin2 5’flanking region (1389bp) was cloned which has strong promoter activity which proved by dual-luciferase reporter assay system.3. Intelectin2 promoter was constructed into PGL3 dual-luciferase reporter vecter. In order to investigate the expression and regulation mechanism of porcine intelectin2, pcDNA3.1-C/EBPa (CREB1, KLF15, PPARy2, SREBP-lc) were transiently transfected into mouse NIH3T3 cells with promoter PGL3 dual-luciferase reporter vecter, respectively. After transfection, we use the dual-luciferase reporter assay system to test the activity of intelectin2 promoter. The results showed that C/EBPa, CREB1, PPARγ2 and KLF15 had no significant effects on the activity of intelectin2 promoter, on the contrary, SREBP-1c could significant inhibit the activity of porcine intelectrin2 promoter.

Related Dissertations

  1. The convergence of mathematics teaching in primary and secondary schools under the new curriculum with the problem representation,G633.6
  2. Studies on Synthesis, Characterization and Properties of Fe-V Co-doped TiO2 Catalyst,O614.411
  3. Characterization of Corn Lectin and Application to Screen of Growth-promoting Bacteria from Corn Rhizosphere,S513
  4. Health-based and Ecological Risk Assessment of Contaminated Sites,X820.4
  5. Preliminary Research on Characterization of Thermophilic Protease Pasey and Helicase Htc16,Q814
  6. Cloning and Expression of Xylanase Gene from Thermobifida Halotolerans YIM 90462~T,Q78
  7. Isolation and Characterization of Carbendazim-Degrading Strains, Cloning and Expression of the mheI Gene,X172
  8. Characterization of an Atrazine-Degrading Strain, Cloning of Key Degrading Related Genes and Construction of a Gene Cluster,X172
  9. Study on the Characteristics of Endoproteases and Changes of Main Nitrogenous Components during Brown Rice Germination,TS210.1
  10. Characterization and Combining Ability of New CMS Lines Improved Using Zhenshan 97 B, an Early Maturity Maintainer, in Indica Rice (Oryza Sativa L.),S511.21
  11. Isolation of Xylanase Producing Bacteria from the Large Intestine of Adult Inner Mongolia Black Sow and Its Enzyme Producing Factors,S816.7
  12. Preparation, Separation of Monascus Pigments and Structural Characterization of Monascus Yellow Pigment,TS264.4
  13. Complexes of two iodine three Allylthiourea together Synthesis and Characterization of cadmium,O627
  14. Substituted phthalocyanine synthesis and molecular self-assembly behavior,O621.3
  15. Study on Preparation of Co-Mo Sulfur-Resistant Shift Catalyst,O643.36
  16. Metallic and semiconducting carbon nanotube density gradient separation,TB383.1
  17. Opera "La Traviata" in the Aria of the Characterization and Teaching,J832
  18. Synthesis、Characterization and Properties of Tetra-carboxyl Tetra(1,4-dithiin) Porphyrazine,O643.36
  19. Template Synthesis of Ag/TiO2, Ni2+/TiO2, SnO2/ZnO Hollow Nano-fibers and Research of Their Photocatalytic Properties,O643.36
  20. Effect of Palmitic Acid on the Fatty Degeneration of Hepatic Cells Induced by Ehanol in Vitro,R575
  21. Preparation and Characterization of Magnetic Molecularly Imprinted Polymer Microsperes,O631.3

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