Cloning and clinical application of telomerase cDNA

本课题的新发现是：1）从猪睾丸中分离纯化出部分端粒酶，聚丙烯酰胺凝胶电泳显示少量蛋白条带，但自端粒酶催化亚基cDNA（hTERT）克隆以来，已有研究者报道了端粒酶催化亚基的克隆（1997年8月），我们放弃了通过蛋白质，纯化，2）hTERTmRNA的表达与端粒酶活性的存在相关; 3）端粒酶阴性的人成纤维细胞经hTERTcDNA转染后表达端粒酶活性，（4）端粒酶阳性的人成纤维细胞端粒长度和增殖寿命均延长，但不能永生化。（5）端粒酶阴性的人成纤维细胞经SV 40 T抗原转化后也表达端粒酶活性。6）这些端粒聚合酶阳性的SV 4 O转化的人成纤维细胞延长了端粒长度和增殖寿命，并且它们永生化。

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.