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| cDNA文库法制备HCV-1诊断芯片探针的应用性研究 | |
| 引用本文： | 孙朝晖,郑文岭,张宝,彭翼飞,马文丽.cDNA文库法制备HCV-1诊断芯片探针的应用性研究[J].生命科学研究,2004,8(3):256-259. |
| 作者姓名： | 孙朝晖 郑文岭 张宝 彭翼飞 马文丽 |
| 作者单位： | 1. 第一军医大学,基因工程研究所,中国,广东,广州,510515;广州军区广州总医院,肿瘤分子生物学研究所,中国,广东,广州,510010 2. 广州军区广州总医院,肿瘤分子生物学研究所,中国,广东,广州,510010 3. 第一军医大学,基因工程研究所,中国,广东,广州,510515 |
| 基金项目： | 国家自然科学基金资助项目(39880032)，广州市重点科技攻关项目(99-Z-022-01) |
| 摘 要： | 探讨利用eDNA文库法制备HCV-1诊断基因芯片探针的可行性．用限制性内切酶Sau3AI消化HCVla及lb全长eDNA，所得的酶切片段72℃补平加A，AT克隆，PCR初步鉴定，并测序．结果显示：HCV两个亚型1a、1b的全长eDNA得到57个大小相对一致(200-1000bp)的片段，平均每个亚型约28个，PCR及序列分析表明，所扩增的片段均属于HCV-1的特异基因，可做为HCV-1诊断基因芯片探针．利用eDNA文库法收集片段是一种快速、简便制备芯片探针的实用方法． |
| 关 键 词： | HCV 分子克隆 微阵列 分子探针 |
| 文章编号： | 1007-7847(2004)03-0256-04 |
| 修稿时间： | 2004年2月3日 |
DNA Microarray Probe Preparation Using cDNA Fragments Library from HCV Genotype 1 |
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| SUN Zhao-hui ,ZHENG Wen-ling ,ZHANG Bao ,PENG Yi-fei ,MA Wen-li.DNA Microarray Probe Preparation Using cDNA Fragments Library from HCV Genotype 1[J].Life Science Research,2004,8(3):256-259. | |
| Authors: | SUN Zhao-hui ZHENG Wen-ling ZHANG Bao PENG Yi-fei MA Wen-li |
| Institution: | SUN Zhao-hui 1,2,ZHENG Wen-ling 2,ZHANG Bao 1,PENG Yi-fei 1,MA Wen-li 1 |
| Abstract: | The technique of cDNA fragments libr ary construction in the preparation of HCV-1probes for clinical diagnostic microarray were studied.The full-length HCV cDNAs of distinct subtypes of 1a and 1b were digested with restriction endonuclease Sau3A I and the resulting fragments were cloned into the pMD18-T vectors.Positive clones were isola ted and identified by sequencing.A t otal of 57different fragments were obtained and sequences analysis showed that all the fragments ranging fr om 200to 1000bp were specific gene fragments of HCV genotype 1,which ca n be efficiently used in microarray p repapration.The method of preparing microarray probes by construction of cDNA fragments library was effective,quick and simple. |
| Keywords: | HCV clone microarray gene probes |
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