Cloning and clinical application of telomerase cDNA

本项目的新发现有：1)从猪睾丸中部分纯化到端粒酶，在丙烯酰胺凝胶电泳上显示出几个蛋白带，但由于其他研究者已经报道了端粒酶催化亚基的cDNA （hTERT）克隆（1997年8月），我们放弃了通过蛋白纯化的克隆项目；2)发现hTERT mRNA的表达与端粒酶活性存在相关。3)转染hTERT cDNA后，端粒酶阴性的人成纤维细胞表达端粒酶活性；4)端粒酶阳性的人成纤维细胞延长了端粒长度和增殖寿命，但没有长生不老；5)转染SV40 t抗原后，端粒酶阴性的人成纤维细胞也表达了端粒酶活性。转染hTERT cDNA后，这些端粒酶阳性的sv40o转化的人成纤维细胞延长了端粒长度和增殖寿命，并实现了长生不老。

New findings of this project are 1) telomerase was partially purified from porcine testis to a few protein bands on acrylamidegel electrophoresis, but, since cDNA (hTERT) cloning of assumed catalytic subunit of telomerase has been reported by other reserchers (Aug. 1 997), we abandoned cloning project through protein, purification, 2) expression of hTERT mRNA was found to be correlated with presence 0f telomerase activity, 3) telomerase-negative human fibroblasts expressed telomerase activity after transfection with hTERT cDNA, 4) these telomerase positive human fibroblasts extended both telomere length and proliferative lifespan, but they did not immortalized, 5) telomerase-negative human fibroblasts transformed with SV40 T-antigen also expressed telomerase activity after. transfection with hTERT cDNA, 6) these telomerase-positive SV4O-transformed human fibroblasts extended both telomere length and proliferative lifespan, and they immortalized.