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Screening of differentially expressed genes in epithelial cells of age-related cataract and normal lens
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 12
- DOI:
- 10.3321/j.issn:1000-2790.2003.12.003
- Key Word:
- 消减杂交法;园翳内障;晶体/细胞学;上皮细胞;差异表达基因
Abstract： 目的: 筛选老年性白内障与正常晶体上皮细胞的差异表达基因.方法: 利用改良消减杂交法获得差异cDNA,利用PCR法扩增其中大片段差异cDNA,将其克隆入质粒载体,通过反向点杂交排除假阳性克隆后,阳性克隆测序与GenBank中的序列进行比较,新序列进行计算机分析.结果: 在文库中随机挑选约600个克隆酶切鉴定,获得≥750 bp片段400个.经反向点杂交,排除假阳性克隆后,共得到阳性克隆150个;测序的50个片段中,有30个已知基因(包括20个功能已知基因、10个序列已知但功能未知基因),3个新序列.新序列中有一个的氨基酸序列与假想的锌指蛋白有45%同源性,可能参与细胞生长调控与代谢.结论: 改良消减杂交法可以快速有效地获得老年性白内障差异表达基因,对稀有基因及新基因的发现与功能鉴定有重要意义.
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