| Part one: The Function of Vsx1 during Embryogenes is of GoldfishVsx1 was proposed to be a regulatory gene related with spatial patterning of eyes of goldfish and diploid-dependent of regulatory mechanism. In order to investigate the function of Vsx1 in the development of Goldfish embryos, and the role in diploid-dependent of regulatory mechanism , we performed a Morpholino antisense Oligos inhibition experiment with Microinjection, and found the change of proteome due to the silence of Vsx1, then investigated the function and the regulatory pattern of Vsx1.We took embryos injected with Morpholino antisense Oligos (ATG-MO) as sample group, and embryos injected with sterile water as control group. Having observed the development of embryos, we found that there are 67. 6 percent embryos of sample group couldn' t continue to develop in the early period of gastrulation. On tne contrary, only 13.5 percent embryos of control group died because of injection or digestion. It indicated that ATG-MO leaded to the silence of Vsx1 in gastrula stage. As we know, early embryo growth mainly depends on parental mRNAs and proteins, after the midblastula transition, the gene group starts to activate, and the gastrulation is the crucial time for organogenesis anlage and the formation of endoderm, mesoderm and ectoderm, therefore, Vsx1 is very possible one of genes which participate in the specification and differentiation of germinal layer in the goldfish embryonic development process. Research reports mainly concentrated on its function for its central nervous system growth and the retina growth. Our research discovered Vsxl is very possible a multi-purpose gene, Vsxl is very possible one of genes which participate in the specification and differentiation of germinal layer during the goldfish embryonic development process.Differential proteome expression analysis was performed by using 2-DE(two dimensional gel electrophoresis) and PDQUEST software, about one hundred spots were visualized, and sixty spots were identified by Mass spectrometry. Nineteen protein spots only expressed in sample group, and eight vice versa; thirteen protein spots were upregulated and twenty protein spots were downregulated in sample group. According to the function of the identified proteins, many kinds of transcription factors were found in the downstream of Vsxl. The proteins such as putative oncoprotein nm23, Cct4 protein and so on were all upregulated in the sample group, It accorded with the previous report and suggested that Vsxl was a transcriptional repressor. It was suggested that Vsxl may indirectly regulate various function genes through these transcription factor; And also some differentially expressed proteins related with signal transduction,cell differentiation, nervous development, these are all possible directly or indirectly regulated by Vsxl. Part two: The Construction and Searches of Differentially Expressed Protein DatabaseComparative proteomics is a major research area in proteomics era, which mainly aims to detect and confirm the different proteins caused by special stimulations between two or more group of samples. By analysis of the difference between samples, in reverse, we can deduce what induce the variety patterns of expression, so it is useful in cancer pre-diagnosis, new drug development and cell regulation research. Hence, more and more Research Workers engage themselves in Comparative proteomics, and as the development of high throughput technology, the data of differentially expressed proteins are increasing day by day. The DEPD(Differentially Expressed Protein Database) is designed to store the output of comparative proteomics studies and provides a publicly available query and analysis platform for data mining. The database contains information about more than 3000 DEPs (differentially expressed proteins)manually extracted from the published literature, including relevant biological, experimental and methodological elements. Mostly, they are related with severe disease, such as lung cancer, breast cancer and liver cancer. The DEPD integrates functional information from the Swiss-Prot/TrEMBL, GO, KEGG and Pfam databases with published studies of DEPs. We set up a user-friendly web interface with tools for querying, and analysis of the results of published comparative proteomics studies. All of the DEPD data can be downloaded freely from the download page of website.Availability: http://protchem.hunnu.edu.cn/depd/... |
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