Cloning And Characterization Of Three Novel CDNAs Expressed In Human Brain, GRY-RBP, LDB1 And BL1

The neural development is one of the major fields in neuroscience. The research of the neural development was accelerated due to introducing molecular biology techniques in the past years. But until now, most knowledges about the neural development are from model organisms, and little is known about the molecular mechanism of human neural development. For the reason, more and more laboratories are involved in the isolation and characterization of the genes expressed in human brain.In order to isolate and clone novel genes related to the development of the human nervous system, we analyzed the ESTs isolated by our lab and the genes related to neural development in model organism. Some ESTs were chosen as the probes for cDNA library screening. For screening the cDNA library quickly and efficiently, an arrayed cDNA library about 3x10~6 individual cDNA clones and an improved PCR-based technique were established. This PCR-based technique features high specificity, high efficiency and less-cost in large-scale cDNA library screening. Our implementation of the technique resulted in the isolation of several full length cDNA clones from the human fetal brain cDNA library. In this paper, the cloning and characterization of 3 full-length cDNAs were reported.A novel human cDNA of 2932 bp has been cloned and named GRY-RBP, which encodes a protein of 623 amino acids with a deduced molecular weight of 69.6 kDa. The deduced protein of GRY-RBP is a RNA binding protein containing three RNA Recognition Motifs (RRMs) and a glycine(G), arginine(R), tyrosine(Y)-rich C-terminus. Computer searches with sequence database revealed high homology to hnRNP R(about 82% identity) and moderate homologies with RRMs of many RRM RNA binding proteins. Using the primers for human GRY-...