Expression, Purification Of HTRT Functional Motifs In E.coli And Preparation Of Antibody Against HTRT Functional Motifs

Linear eukaryotic chromosome are capped by tandem repeats of simple sequence,G-rich and oriented 5' -3' toward the chromosome ends. These termini,called telomere,protects the chromosome ends from degradation,recombination,and complete replication of the chromosome ends . For telomere shorten with each round of cell division,these will lead to cell senescence. Human telomeric sequence is estimated as 5-20 kb .which consists of regular repeat (ITAGGG).Telomerase losts 50-200 bp at every somatic cell division. Eventually,if unckecked,progressive telomere erosion leads to cellular death.Normal cultured cells have limited capacity of replication because of telomere shorten,whereas in tumor cells and other immortal cells can maintain telomere length,which indicates telomere sequence is re-added onto chromosome end. It is now clearly that the enzyme telomerase activated can add sequence to the end of telomere. Telomerase is a sepecial RNA dependent DNA polymerase,which maintains telomere length and prevents cell senescence,and contains telomerase RNA component and protein components.. It appears to be selectively expressed in tumors versus normal cells,and this enzyme is considered as a good biomarker for cancer,whereas telomerase activation mechanism has not been clearly interpreted.Telomerase RNA component and Telomerase protein components,has been isolated .cloned and sequenced from human,which highly propels telomerase studies in the fields of telomerase activation mechanism,structure,inhibitorand others. Telomerase -associated Protein (TP1) shows specifically interact with telomerase RXA subunit,and these components are not required for in vitrotelomerase activity. The key protein component is telomerase reverse transcriptase (hTRT) catalytic subunit.a limiting factor for its activity,which contains seven reverse transcriptase motifs and sepcific region with conserved sequence termed 1