Dissertation > Excellent graduate degree dissertation topics show

ES Mice Production and Expression of Surfactant Protein B mRNA

Author: WuXingLong
Tutor: LiXiangYun
School: Agricultural University of Hebei
Course: Animal Genetic Breeding and Reproduction
Keywords: ES mice Respiratory failure syndrome Pulmonary surfactant protein B Tetraploid embryos ECCD-1 Microinjection
CLC: S865.13
Type: Master's thesis
Year: 2011
Downloads: 13
Quote: 0
Read: Download Dissertation

Abstract

ES mice belonging to clone mice, the best model animal for the study of human disease. Use of the tetraploid embryos compensation technology can be completely developed from mouse ES cells, ES mice. Although the the tetraploid embryos compensation technology we recognized, but with lower the ES mice efficiency of this technology to prepare, many influencing factors. The number of passages in production methods and genetic background of tetraploid embryos cultured in vitro environment, ES cells are important factors affecting ES mice. Tetraploid embryo developmental stage is also an important factor, about the tetraploid embryo development stage ES mice prepared yet. Freshmen viable ES mice (brief breathing phenomenon) is not able to develop to adulthood, a considerable part of performance progressive dyspnea and cyanosis signs of respiratory failure immediately after birth, their life will not be more than one hours, lung pathology performance as a thickening of the alveolar walls and alveolar atelectasis, and this respiratory failure levy is the major diseases leading to death of the newborn ES mice. Newborn ES mice died of respiratory failure almost no expression of SP-B protein, even if the normal appearance of adult ES mice SP-B protein levels were also very low. SP-B gene is likely to be to ensure that the newborn cloned animals survived to adulthood key genes. 2 - cell embryos the electrofusion prepared tetraploid embryos, microinjection method, ES cells were injected into 1 - cells - cells tetraploid embryos in the blastocyst developmental stages. The ES cells were B6D2F1 × 129/Sv hybrid lines and inbred C57BL/6J, the ES mice embryo transfer and caesarean section. The results show that the hybrid ES cells were injected tetraploid blastocysts obtained 40 ES mice efficiency is significantly higher than inbred ES cells as well as other developmental stage tetraploid embryos, ES mice and B6D2F1 × 129/Sv hybrid mice coat consistent with normal reproductive capacity. Experiments, we also studied the E-type cadherin antibody (ECCD-1) of the embryos in chimera ECCD-1 deal with the 8-cell embryo injection of ES cell method. The results show that, ECCD-1 prevents the embryo splitting the connection between the ball, to postpone the densification process, but does not affect the formation of the blastocyst ICM. ECCD-1 for 8 - cell embryos injection of ES cells can be completely developed from mouse ES cells. 14 died of respiratory failure in the newborn of ES mouse and 14 newborn lung tissue of normal mice (same genetic background) were taken, and RNA was extracted. Semi-quantitative PCR method for detection of SP-B mRNA expression level, the results show that the newborn ES mice died of respiratory failure in the lung tissue of SP-B mRNA decreased expression. These results indicate that, neonatal respiratory failure levy cloned animals is likely to be caused by the abnormal expression of the SP-B.

Related Dissertations

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Hunting,domestication of wild animals > A variety of wild animal domestication > Animals used for experiments > Mice
© 2012 www.DissertationTopic.Net Mobile