基因BC023882发育学表达模式、亚细胞定位及其对细胞周期的影响
发布时间：2018-08-26 16:54
【摘要】： 研究背景 前期课题“高密度cDNA基因芯片筛选小鼠脑发育、损伤相关基因的研究”利用含小鼠发育文库cDNA 7680条的高密度基因芯片,对发育和损伤模型中表达差异的基因进行筛选。结果发现发育过程中高表达基因560个,其中过度表达有34个。这些基因功能广泛,涉及转录调节、信号转导、细胞代谢、免疫调节、细胞骨架、DNA复制修复、细胞粘附等17类。其中一个功能未知的过度表达基因BC023882引人注目,它在胚胎发育和生后早期小鼠的脑组织存在高表达,其中P0 d时相点Cy5/Cy3比值(Ratio值)为7.6038,远远高于2.0000的基因表达差异筛选标准。提示基因BC023882可能在小鼠早期胚胎中枢神经系统的发育过程中起到重要作用。研究方法与结果 1.基因BC023882的发育学表达模式及脑区定位。 (1)半定量RT-PCR检测基因BC203882在发育小鼠各个阶段的表达情况。分别取小鼠胚胎期时相点(E14、E19 d)及生后发育时相点(P1、P7、P14、P30、P60 d)的小鼠组织。其中胚胎期组织包括脑(brain)和胚体(body),而生后组织包括脑、肝、肾、心、肺和小肠。分别提取上述组织的总RAN,利用RT-PCR技术检测基因023882在上述组织中的表达。我们通过RT-PCR实验观察到基因BC023882的mRNA呈阶段性地表达于脑组织,P1 d达到高峰,P60 d表达消失;其mRNA也少量表达于胚体组织,具体定位有待进一步实验确定;另外P1 d肝组织也有微弱表达,随后迅速消失,而肾脏、肺、小肠和心脏等组织均未检测到该基因表达。 (2)用原位杂交技术检测基因BC023882的mRNA在不同脑区中的定位。根据基因BC023882的特异性序列设计RT-PCR引物,扩增后构建在含T7启动子的pcDNA3.1(-)载体中,标记cRNA探针。利用该基因探针对各实验组脑组织进行原位杂交。通过组织原位杂交实验观察:基因BC023882的mRNA的最早出现在E14 d的胚胎脑组织大脑皮层,包括运动皮层、躯体感觉皮层、扣带皮质和梨状皮质;P1 d时杂交阳性细胞出现范围更广,如大脑皮层,内囊、胼胝体、视前区诸核团等等;杂交信号
[Abstract]:Background "screening of mouse brain development and damage related genes by high-density cDNA gene chip" using a high-density gene chip containing cDNA 7680 in mouse development library. The differentially expressed genes in developmental and injury models were screened. The results showed that there were 560 overexpression genes, 34 of which were overexpressed. These genes have a wide range of functions, including transcriptional regulation, signal transduction, cellular metabolism, immunomodulation, cytoskeleton DNA replication and repair, cell adhesion, and so on. One of the unknown overexpression genes, BC023882, was highly expressed in the brain of embryonic and early postnatal mice. The P0 day phase point Cy5/Cy3 ratio (Ratio) was 7.6038, which was much higher than the differential screening criterion of gene expression of 2.0000. The results suggest that gene BC023882 may play an important role in the development of central nervous system in early embryonic mice. Methods and results 1. (1) Semi-quantitative RT-PCR was used to detect the expression of gene BC203882 in various stages of development mice. The tissues of mouse embryonic phase (E14, E19 d) and postnatal developmental phase (P1P7, P14, P30, P60 d) were taken respectively. Embryonic tissues include brain (brain) and embryonic body (body), while postnatal tissues include brain, liver, kidney, heart, lung and small intestine. Total RAN, was extracted from the above tissues to detect the expression of gene 023882 by RT-PCR technique. Through RT-PCR experiments, we observed that the mRNA of gene BC023882 was periodically expressed at the peak of P1d in brain tissue and disappeared at P60 day, and its mRNA was also expressed in embryoid tissue. The specific location of mRNA was to be determined by further experiments. In addition, there was also a weak expression in P1d liver tissue, and then disappeared rapidly, while the kidney, lung, No expression of the gene was detected in small intestine and heart. (2) in situ hybridization technique was used to detect the localization of mRNA of gene BC023882 in different brain regions. According to the specific sequence of gene BC023882, RT-PCR primers were designed and amplified into pcDNA3.1 (-) vector containing T7 promoter and labeled with cRNA probe. The gene probe was used for in situ hybridization of brain tissue in each experimental group. By tissue in situ hybridization, the mRNA of gene BC023882 was first found in the cerebral cortex of embryonic brain on day 14, including motor cortex, somatosensory cortex, cingulate cortex and piriform cortex. Such as cerebral cortex, internal capsule, corpus callosum, preoptic nuclei, etc.; hybridization signals
【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329
[Abstract]:Background "screening of mouse brain development and damage related genes by high-density cDNA gene chip" using a high-density gene chip containing cDNA 7680 in mouse development library. The differentially expressed genes in developmental and injury models were screened. The results showed that there were 560 overexpression genes, 34 of which were overexpressed. These genes have a wide range of functions, including transcriptional regulation, signal transduction, cellular metabolism, immunomodulation, cytoskeleton DNA replication and repair, cell adhesion, and so on. One of the unknown overexpression genes, BC023882, was highly expressed in the brain of embryonic and early postnatal mice. The P0 day phase point Cy5/Cy3 ratio (Ratio) was 7.6038, which was much higher than the differential screening criterion of gene expression of 2.0000. The results suggest that gene BC023882 may play an important role in the development of central nervous system in early embryonic mice. Methods and results 1. (1) Semi-quantitative RT-PCR was used to detect the expression of gene BC203882 in various stages of development mice. The tissues of mouse embryonic phase (E14, E19 d) and postnatal developmental phase (P1P7, P14, P30, P60 d) were taken respectively. Embryonic tissues include brain (brain) and embryonic body (body), while postnatal tissues include brain, liver, kidney, heart, lung and small intestine. Total RAN, was extracted from the above tissues to detect the expression of gene 023882 by RT-PCR technique. Through RT-PCR experiments, we observed that the mRNA of gene BC023882 was periodically expressed at the peak of P1d in brain tissue and disappeared at P60 day, and its mRNA was also expressed in embryoid tissue. The specific location of mRNA was to be determined by further experiments. In addition, there was also a weak expression in P1d liver tissue, and then disappeared rapidly, while the kidney, lung, No expression of the gene was detected in small intestine and heart. (2) in situ hybridization technique was used to detect the localization of mRNA of gene BC023882 in different brain regions. According to the specific sequence of gene BC023882, RT-PCR primers were designed and amplified into pcDNA3.1 (-) vector containing T7 promoter and labeled with cRNA probe. The gene probe was used for in situ hybridization of brain tissue in each experimental group. By tissue in situ hybridization, the mRNA of gene BC023882 was first found in the cerebral cortex of embryonic brain on day 14, including motor cortex, somatosensory cortex, cingulate cortex and piriform cortex. Such as cerebral cortex, internal capsule, corpus callosum, preoptic nuclei, etc.; hybridization signals
【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329
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