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| 一种用少量标本快速构建老年性白内障消减cDNA文库的方法 | |
| 引用本文： | 刘燕,周健,刘新平,药立波,王吉村,苏景宽,惠延年.一种用少量标本快速构建老年性白内障消减cDNA文库的方法[J].生物化学与生物物理进展,2002,29(6):952-954. |
| 作者姓名： | 刘燕 周健 刘新平 药立波 王吉村 苏景宽 惠延年 |
| 作者单位： | 1. 第四军医大学西京医院眼科,西安,710032 2. 第四军医大学生物化学与分子生物学教研室,西安,710032 3. 第四军医大学唐都医院医教部,西安,710032 |
| 基金项目： | 国家自然科学基金资助项目（30000185）. |
| 摘 要： | 为从少量标本中获得含较多大片段的、高质量的老年性白内障消减cDNA文库，利用磁珠分离、生物素标记的改良消减杂交法获得差异cDNA，利用选择性PCR法扩增其中大片段差异cDNA，从而成功构建老年性白内障消减cDNA文库.在文库中随机挑取的22个克隆中，1 000 bp以上的片段有7个，占31.8%，750 bp以上有15个，占68.2%.所得cDNA片段较大，可以满足下一步研究需要.改良消减杂交法结合选择性PCR法可以从少量标本中快速有效地获得大片段高质量的消减cDNA文库. |
| 关 键 词： | 改良消减杂交法，选择性PCR法，老年性白内障，消减cDNA文库 |
| 收稿时间： | 2002/5/27 0:00:00 |
| 修稿时间： | 2002年5月27日 |
A Rapid Procedure for The Construction of Subtractive cDNA Library of Age-related Cataract From Small Amounts of Sample |
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| LIU Yan,ZHOU Jian,LIU Xin-Ping,YAO Li-Bo,WANG Ji-Cun,SU Jing-Kuan and HUI Yan-Nian.A Rapid Procedure for The Construction of Subtractive cDNA Library of Age-related Cataract From Small Amounts of Sample[J].Progress In Biochemistry and Biophysics,2002,29(6):952-954. | |
| Authors: | LIU Yan ZHOU Jian LIU Xin-Ping YAO Li-Bo WANG Ji-Cun SU Jing-Kuan and HUI Yan-Nian |
| Affiliation: | Department of Ophthalmology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032,China;Department of Ophthalmology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032,China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032,China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032,China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032,China;Department of Science Reserch, Fourth Military Medical University, Xi'an 710032,China;Department of Ophthalmology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032,China |
| Abstract: | In order to obtain a quality subtractive cDNA library of age-related cataract which contains more large fragments from small amounts of sample, an improved subtractive hybridization which using magnetic beads isolation and biotin labelling was used to obtain the subtractive cDNA. Then the large fragments of the subtractive cDNA were amplified by using selective PCR method, so the subtrative cDNA library of age-related cataract was successfully constructed. 22 clones of the library were selected at random and were identified by the restriction enzymes digestion. 7 fragments were larger than 1 000 bp,which is 31.8% of the totals, 15 fragments were larger than 750 bp, which is 68.2% of the totals. The cDNA fragments are larger and can be used in further study. A subtractive cDNA library with high quality and more large fragments can be constructed rapidly and effectively by improved subtractive hybridization and selective PCR method from small amounts of sample. |
| Keywords: | inproved subtractive hybridization selective PCR age-related cataract subtractive cDNA library |
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