The Class Of Sulfur Redox Protein Of Cdna Cloning And Preliminary Functional Studies

Human brain plays a crucial role in the life of individual, from embryonic development to growth, maturation and aging. Human genome project which aims to decode the whole nucleotide sequence of 3×10~9 bp of human genome brings new opportunity for developmental neuroscience. Expressed genes are also most abundant in brain, therefore it becomes an important resource for the isolation of expressed sequences tags (ESTs) and cloning of novel human genes. Many expressed sequences in brain have been successfully located on human chromosomes, which provide an solid base for the structural and functional study of genes expressed in brain. An international brain project which is one of the most important parts of human genome project has widely developed. The cloning and study of genes related to the disease and development of central nervous system have become not only the research hot spot of modern genomics , but also the keystone of molecular genetics and developmental biology.The main research field of our lab was focused on the isolation and cloning of genes related to the development and disease of human brain. Human fetal brain tissues at different developmental stages(13 week cerebrum and 33 week cerebrum) and also human glioma cell line BT325 before and after induction of differentiation with sodium butyrate were subject to mRNA differential display analysis (DDRT-PCR). The isolated ESTs with different expression patterns were cloned into pBlue-Script and pUC19 vectors and then were sequenced. Sequence similarity search was conducted to analyze the ESTs by using Blast algorithm. Some ESTs were chosen as the probes for cDNA library screening. In order to screen the cDNA library quickly and efficiently, an arrayed human fetal brain cDNA library about 3×10~6 individual cDNA clones were prepared and an improved PCR-based technique were established. The execution of the methods during my thesis work resulted in the isolation and cloning of two novel, full length cDNA clones: hTRXL and CacyBP. Their identification and preliminary functional analysis were carried out.