Retrovirus Vector-Mediated Bystin Gene Expression And Function Research Of Bystin

Retrovirus vector-mediated Bystin gene expression and function research of BystinBystin is an important molecule which consists of 306 amino acid residues and the molecular weight is about 35kD. It was first found by Fukuda et al with yeast two-hybrid assay in 1998. Bystin involved in the initial attachment of blastocysts to the uterus in humans. Our previous experiments revealed that the expression level of Bystin was significantly increased in the substantia nigra following 6-OHDA lesioning and other lesion. Furthermore, double-labeling immunofluorescence showed that Bystin-positive cells were primarily GFAP-labeled astrocytes. These Bystin-positive astrocytes had morphological characteristics of activated astrocytes, it suggests that Bystin is probably involved in neurodegeneration or neuroregeneration, but its exact function in central nervous systerm is unknown. So we established an expression model to study the function of Bystin.Recently, some groups demonstrate that with the recombinant retrovirus vector-mediated gene tranfer, several models of major neurodegenerative diseases, such as Parkinson's Huntington's or Alzheimer's disease can be made.Retrovirus vectors can direct gene transfer to selected subregions or subsets of neurons in the brain. Otherwise, the current retroviral vectors provide a unique opportunity to apply functional genomic techniques to the discovery of new drug targets in a broad spectrum of cell types. Introduction of the libraries of genetic effectors into cells using retroviral vectors yields a population of individually mutagenized cells. Those cells with a new phenotype owing to expression of a specific genetic effector can then be selected and isolated. By defining the interactions of these effectors with other proteins in the cell, potential targets for the development of therapeutic agents can be identified. These successful case of retroviral vector based gene transfer can be used for reference in function research of Bystin.With the retroviral vector pLXSN-Bystin,we successfully direct Bystin gene transfer to primary E14 neural cells of rat. With studies of subcelular localization,we found that Bystin was co-localized with p-tubulin,it suggests Bystin protein might be a microtubule-associated protein in astrocytes. Moreover,using pcDNA3,1-Bystin vector to transfect COS-1 cells,Bystin can also be found in the nucleus of COS-1. simultaneously,apoptosis is induced in some cells,and a majority of these apoptotic cells are not Bystin-positive. These results provide significative data to improve conditions of Bystin expression in vitro or in vivo,and give the basis for further study .