Expression of DNA sequences containing neuron specific enolase gene in Escherichia coli.
There is evidence that the gene for gamma-gamma enolase (neuron specific enolase, NSE) is regulated during cell differentiation and development, conserved in a variety of organisms and contains mRNA destabilizing sequences. In order to investigate further the mechanisms of these processes and to obtain large quantity of this protein, the NSE gene was isolated from neuroblastoma cells and cloned in E. coli using standard molecular biology techniques. The NSE gene expression was studied and the expressed protein (recombinant NSE) was characterized extensively. The recombinant NSE behaves like parental NSE in antisera specificity, resistance for chaotropic agents like urea, thermal stability at higher temperatures etc. The physical parameters like secondary structure, hydrophilicity, antigenic index and flexibility of the expressed protein were studied. The results of the present investigation collectively form the basis for initial investigations of how the expression of NSE gene is regulated. This is the first report where the recombinant NSE gene has been characterized so extensively.
Dutta
S K
SK
Department of Botany, Howard University, Washington, DC 20059.
Bhattacharyya
N
N
Parui
R
R
Verma
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002460:Cell Line; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006822:Hybrid Cells / Q000201:enzymology; D007700:Kinetics; D009474:Neurons / Q000201:enzymology*; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D000596:Amino Acids; D000939:Epitopes; D011994:Recombinant Proteins; D004247:DNA; D010751:Phosphopyruvate Hydratase