Purification, identification of human telomerase and its protein subunits

To purify human telomerase complex and analyse its protein subunit components, an affinity-purification protocol using human telomere repeat sequence as affinity ligand was designed and carried out to purify HeLa cell telomerase. A DIG gel shift protocol was performed to identify the purified component. SDS-PAGE was used to analyze protein subunits of purified component. The component with telomerase activity was obtained from elution fluid. A single band on native gel and a special DIG gel shif band on X-ray film were observed. Four protein subunits were observed using SDS-PAGE. Their relative molecule mass are 220 ku, 212 ku, 116 ku and 43 ku, respectively. Human telomerase complex and its 4 protein subunits were obtained.