人类YAC库PCR三维筛选体系的建立及质量考核

The Establishment of a Three Dimensional PCR Screening System and Quality Evaluation of Human YAC Library

为了在知道某个区域、位点、基因或ＤＮＡ片段的部分信息后，能从ＣＥＰＨＹＡＣ库中筛选出与其对应的ＹＡＣ克隆，为进一步研究奠定基础，需要建立一个筛选体系。本文概述了这一筛选体系的建立过程。随后，用两对与已知基因对应的引物进行了筛选验证工作，证明了这一体系的可用性，同时提出了以后筛选的途径，即首先筛选ＹＡＣ库的ＭｅｇａＹＡＣ部分，并以５块板为一组进行筛选。另外，运用荧光原位杂交技术（ＦＢＨ），对ＣＥＰＨＹＡＣ库的质量及其第一代人类基因组物理图谱进行了考察。我们取２６个ＹＡＣ克隆进行ＦＩＳＨ定位，结果其中嵌合体１３个，占５０％。定位错误的克隆有６个，占２３％。非嵌合体且定位正确的共９个，占３５％。

In order to screen out YAC clones in the CEPH YAC library related to a definite gene,DNA fragment or locus,we need to establish a screening system.This paper includes the establishment of this system and the evaluation of its feasibility.According to the experiments,we suggest that it is better to screen the Mega YAC part of the library at first and put five culture plates as a group.Meanwhile,FISH experiments to assess the quality of CEPH YAC library and the first generation physical map of human genome,Were conducted FISH experiments on 26 YAC clones were corried out.There were 13 clones that were chimeric(about 50%),6 clones that were mapped by mistake(about 23%).So,there were only 9 clones that were mapped correctly and were not chimeric(about 35%).