Dissertation > Excellent graduate degree dissertation topics show

The Construction of cDNA Library from Cerebrum of Elaphodus Cephalophus and Study of Neurotransmitter Transporter Gene

Author: TangWenWen
Tutor: CaoXiangRong
School: Nanjing Normal University
Course: Cell Biology
Keywords: Elaphodus cephalophus Cerebrum cDNA library Neurotransmitter transporter gene Southern blot
CLC: Q78
Type: Master's thesis
Year: 2006
Downloads: 47
Quote: 0
Read: Download Dissertation

Abstract


The Cerebrum cDNA Library from tufted deer (Elaphodus cephalophus) was constructed by using SMART technique. Total RNA was extracted from cerebrum tissue of tufted deer, and poly (A)~+ RNA was purified further by Oligotex mRNA Kit. CDS III/ 3’PCR Primer primed the reverse transcription. The double-strand cDNA was amplified by LD- PCR(long distance PCR) and digested by Sfi I. After cDNA size fractioned by CHROMA SPIN- 400, fragments over 500 bp were collected and ligated with λTripIEx2 vector. After packaged in vitro , the full- long cDNA library of cerebrum tissue was constructed successfully. The library contains 5.1×10~5 clones and the amplified library has a titer of 1.5×10~9 pfu/ml, in which 85% clones are recombinant and the average sizes of inserted cDNAs are about 1.0 kb. These results show that the Cerebrum cDNA Library is qualified for screeening genes. A full- length rig (rat insulinoma gene) gene including 5’ and 3’untranslated regions was isolated from the cDNA library . Sequence analysis shows that this 541 bp cDNA spans an open reading frame from nucleotide 43 to 477, encoding a 51- aminoacid rig protein.In order to get the sex-linked gene, we sequenced and obtained 82 ESTs from testis cDNA Library of tufted deer, 27 genes are 3’-sequence and 55 sequences are 5’-sequence. After homologous comparison in the GenBank with the sequence analysing software BLAST, 4 genes are new and 8 genes are repeated, 3 sequences are located on the X chromosome.The genomic DNAs were extracted from tufted deer and were digested by Hind III respectively. The digested DNAs were transferred on the nylon membrance and hybridization was done between the nylon membrance and the Neurotransmitter transporter gene probe, which was labelled by using PCR DIG labeling Mix. The results of southern blotting showed that the Neurotransmitter transporter gene has two obvious bands in the genome of tufted deer. In order to study the expression of the Neurotransmitter transporter in tufted deer, total RNA was extracted from 8 tissues of

Related Dissertations

  1. Analysis of Infrared Spectral Features and Identification of Al-responsive Genes in Small Black Soybean with Aluminum Stress,S529
  2. Molecular Cloning, Prokaryotic Expression and Characterization of a Novel Trypsin Inhibitor in the Frog, Lepidobatrachus Laevis,Q78
  3. Molecular Cloning and Analysis of Expansin Gene from Heterodera Glycines and Evaluation of Soybean Germpalsms Resistance,S565.1
  4. Construction and Primary Analysis of Differentially Expressed cDNA Library of Alfalfa by Using Suppression Subtractive Hybridization under the High Temperature,S541.9
  5. Construction and Analysis of a Subtractive cDNA Library on Early Embryonic Development Related Genes in Ducks,S834
  6. Cloning and Expression of Novel β-Glucosidase Genes from Rhizopus Stolonifer Var. Reflexus,Q78
  7. Construction of a Full-length cDNA Library and Transcriptome Analysis during Larix Somatic Embryogenesis,Q943.2
  8. Molecular Cloning and Characterization of a New Acetylcholinesterase Gene Bx-ace-2 from Pine Wood Nematode Bursaphelenchus Xylophilus and It’s RNA Interference,S763.3
  9. Construction of the cDNA Library from Stem Cambium Tissue of Tension Wood about Populus Simonii × P.nigra, and Relative Gene Expression Analysis,S792.11
  10. Construction and Analysis of an Inducible Insect Resistant cDNA Library and Its Expressed Sequence Tags from Jasmonic Acid-treated Larix Gmelinii Needles,S791.222
  11. Bioinformatics Analysis on ESTs Sequences from Wheat Stripe Rust Resistance SSH cDNA Library,S512.19
  12. Construction and Analysis of the Subtracted cDNA Library from Hyriopsis Cumingii Liver,S917.4
  13. Construction of cDNA Library and SSH Library of Steinernema Feltiae Infective Juveniles and Genes Expression Profile Analysis,S476.15
  14. Construction and Analysis of Suppression Subtractive Hybridization cDNA Library of Cucumber Induced by Fusarium Oxysporum,S642.2
  15. The Effect of Nicotine Exposure to the Expression of APP and Presenilin 1 in the Cerebrum of Adult Kunming Mice,R749.16
  16. Construction and Sequence Analysis of SMART cDNA Library from Ammopiptanthus Mongolicus under Drought Stress,Q943
  17. The observation of mifepristone on the labor induction of term pregnant rats and its effects on the cerebral tissue of mother rats and their offsprings,R719.31
  18. Cloning, Expression and Antigenicity Analysis of a cDNA Encoding p46 kDa Antigen from Newborn Larvae of Trichinella Spiralis,Q78
  19. Proteomic Analysis of Insecticide Resistance of the White-Backed Planthopper, Sogatella Furcifera (Horvath) (Homoptera: Delphacidae),S435.112.3
  20. Microdissection of H. villosa 6VS conferring resistance to wheat powdery mildew and cloning of disease resistance related genes,S435.121
  21. Cloning of Sex-linked Gene of the Tufted Deer and Identification of Its Sex Chromosomes,Q78

CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
© 2012 www.DissertationTopic.Net  Mobile