【摘要】 目的筛选并鉴定人端粒酶逆转录酶(hTERT)功能区表位。方法应用差减筛选法,以抗hTERT多克隆抗体为靶筛选噬菌体12肽库,通过夹心ELISA、抗hTERT多抗特异性阻断实验、竞争抑制实验鉴定阳性噬菌体克隆并测序。结果经3轮筛选,从随机挑取的24个噬菌体克隆中有13个克隆特异地与抗hTERT多抗结合,而不与正常小鼠IgG结合,其中11个克隆的氨基酸序列富含组氨酸(最高达41.6%)及亲水氨基酸(最高达91.67%)。结论得到11个序列不同的hTERT表位,为研制针对hTERT的小分子抑制剂提供了实验依据。更多还原

【Abstract】 Objective To identify and characterize the epitopes of human telomerase reverse transcriptase (hTERT). Methods A random phage displayed dodecapeptide library was screened with anti-hTERT antibody. The selected phage clones were identified by sandwich enzyme-linked immunosorbent assay, anti-hTERT antibody blocking assay and competitive inhibition assay. Results After 3 rounds of screening, 13 of 24 randomly selected phage clones were identified as positive clones that could specifically bind to anti-hTERT antibody but not to normal mouse IgG. Amino acid sequences deduced from DNA sequences showed 11 different sequences with high percentage of histone and hydrophilic amino acids. Conclusion Eleven epitopes have been obtained that can effectively mimic hTERT, which will be useful for the research of small-molecule inhibitor targeting at hTERT.更多还原