Cloning of telomere-associated protein T-STAR and the interaction between T-STAR and hTERT
- VernacularTitle:人端粒相关蛋白T-STAR的克隆及其与端粒酶催化亚单位hTERT的相互作用研究
- Author:
Ping ZHOU
;
Dianchun FANG
;
Gaoping MAO
- Publication Type:Journal Article
- Keywords:
telomerase;
telomere-binding proteins;
two-hybrid system techniques
- From:
Medical Journal of Chinese People's Liberation Army
1983;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone the telomere-associated protein T-STAR and study the relationship between T-STAR and the telomerase catalyzed subunit hTERT in mammalian cells. Method The expression vector pGBKT7-hTERT was constructed and acted as a decoy in cDNA library screened by yeast two-hybrid technology. Recombinant vectors pVP16-T-STAR and pM-hTERT were constructed and co-transfected with report gene CAT into SGC-7901 cells with liposome. pM-53+pVP16-T and pM3-VP16 were introduced as positive controls, pM-53+pVP16-CP as negative control, and pM-hTERT+pVP16, pM+pVP16-T-STAR and pM+pVP16 as crossing negative controls. The expression of CAT was assayed by ELISA. Results The eukaryotic expression vector pGBKT7-hTERT was successfully constructed. One positive clone achieved by cDNA library screening was sequenced and compared with the isogenous sequences in GenBank by Blast software via Internet. As a result, T-STAR, a recorded cDNA sequence was obtained. The recombinant vectors of pVP16-T-STAR and pM-hTERT were constructed successfully and co-transformed into gastric cancer cells SGC-7901. The OD value of reported gene CAT was 0.258, which was significantly higher than that of the negative and crossing negative controls (0.002-0.015). It revealed that T-STAR interacted with hTERT in the mammalian cells. Conclusions T-STAR interacts with hTERT in the gastric cancer cells. T-STAR may be a new member of telomere-associated protein, and it participates in the regulation of telomerase through hTERT.
