Dissertation > Excellent graduate degree dissertation topics show

Creation and Identification of DFNA64 Mouse Models

Author: LuYu
Tutor: YuanHuiJun
School: PLA Postgraduate Medical School
Course: Department of Otolaryngology - Head and Neck Surgery
Keywords: Mice Transgenic Gene knock Autosomal dominant non- syndromic deafness
CLC: R764.43
Type: Master's thesis
Year: 2011
Downloads: 44
Quote: 0
Read: Download Dissertation

Abstract

The DFNA64 the research group in an autosomal dominant inheritance of non-syndromic deafness pedigrees positioning deafness genes, known by positioning the region gene by direct sequencing find the DIABLO gene c.377C gt co-segregated with the deafness phenotype; heterozygous mutation. DIABLO is very important in the inner mitochondrial membrane cell apoptosis-promoting protein, our group studies suggest p.S126L mutant is more sensitive to external stimuli, can cause mitochondrial membrane potential chronic injury. Further research is needed to replicate in animal models with human clinical performance, and thus validation and analysis of mutant DIABLO damage to the inner ear hair cells, thus revealing the disease-causing gene mutations cause deafness mechanism mechanism for the prevention and treatment of the possibility to lay foundation. A. DFNA64 transfected gene in mice model identified in this study using mammalian efficient expression vector pcDNA3.1 () construct contains the CMV promoter and artificial build intron, cDNA coding sequence of the target gene, the polyadenylation signal components of the transfected gene vectors, through the original nuclear DNA microscopic injection transfected gene technology will construct the outer source transfected gene vector DNA fragment injected into CDl far cross Department of mice homozygous for the former nucleus, respectively, to establish and has been integrated with the mutant and wild-type Diablo gene sequence the DFNA64 transgenic mouse model; to PCR identification mice genotype; primary positive transgenic mice selfing create multiple transgenic lines homozygous transgenic mice; real-time PCR detection Diablo gene expression in the nervous system tissue amount confirmed by the overexpression of the Diablo genes in the DFNA64 transfected gene mice. . DFNA64 gene knockin mouse model to establish this study the use of fixed-point the DFNA64 gene homologous recombination and the establishment of ES cell-based gene targeting technology knockin mice model placed in the 129 strain ES cell genome and human DFNA64 mutation the source p.S124L mutant, chimeric mice. The design of the carrier with homologous sequences of one long and one short paragraphs purpose, the homologous sequence inserted between the mutant sequence and a loxP site anchored positive selection screening neo gene, and the linearized vector DNA is electrically transferred to the ES cells, ES cells integrated into the ES cell genome sequences, by site-directed homologous recombination joint use of antibiotics G418 and long-range PCR screening, Excluding integration and random insertion of the vector sequence screened positive ES cells were microinjected into host blastocysts in vitro the pseudopregnant mice transplanted into synchronization After incubation, develop into chimeric mice. The DFNA64 the establishment of a mouse model for DIABLO gene function, deafness mechanisms, prior to the onset of prevention, drug screening and subsequent research platform.

Related Dissertations

CLC: > Medicine, health > Otorhinolaryngology > Otology,ear disease > Ear nervous system diseases > Deaf
© 2012 www.DissertationTopic.Net Mobile