摘要
分离和克隆ＹＡＣ插入片段的末端顺序是构建ＹＡＣ重叠群的重要手段之一，我们采用Ａｌｕ载体（Ａｌｕ－ｖｅｃｔｏｒｅｔｔｅ）ＰＣＲ方法成功地从含人淀粉样蛋白前体（ＡＰＰ）基因的法国人类多态研究中心（ＣＥＰＨ，Ｃｅｎｔｒｅｄ＇ＥｔｕｄｅｄｕＰｏｌｙｍｏｒｐｈｉｓｍｅＨｕｍａｎ）ＹＡＣ克隆５９９Ｇ１１的未端分离到一个０．５８ｋｂ的单拷贝片段。测序后经核普酸顺序检索分析，证明这是一个新的ＳＴＳ顺序。用这个片段作探针，在英国肿瘤研究基金会（ＩＣＲＦ）的ＹＡＣ库中筛选到一个新的ＹＡＣ克隆，证明这是获得ｃｏｎｔｉｇ的有效而快捷的方法。
The important advance in chromosome walking has been the introduetion of yeast artificialchromcaome(YAC).To construct a YAC contig,cloning the end of YAC insert is a important but ratelimited step.Many methods have been developed.Presently we have cloned two end fragments from twoYACs,which are from CEPH YAC library.Onc is 599g 11 located in 21q11.2,the amyloid proteinprecursor(APP)region,and another is 828b09 which is located 17p,The left end of 599911 and right endof 828b09 have been cloned.The later was verified containing repetitive sequence after hybridization withhuman genomic DNA,The former is a single copy sequence and has been seq uenced.The database has beenscanned.There is no similar sequence in it. So the left end of 599g 11 is a new STS of APP region.screening the ICRF YAC Iibrary with the end,one YAC clone has been picked up.
出处
《高技术通讯》
CAS
CSCD
1995年第3期40-43,共4页
Chinese High Technology Letters
基金
863计划及国家自然科学基金
