牛酵母人工染色体（YAC）文库的构建。

Construction of a bovine yeast artificial chromosome (YAC) library.

作者信息

Takeda H, Yamakuchi H, Ihara N, Hara K, Watanabe T, Sugimoto Y, Oshiro T, Kishine H, Kano Y, Kohno K

机构信息

Shirakawa Institute of Animal Genetics, Fukushima, Japan.

出版信息

Anim Genet. 1998 Jun;29(3):216-9. doi: 10.1046/j.1365-2052.1998.00325.x.

DOI:10.1046/j.1365-2052.1998.00325.x

PMID:9720181

Abstract

We have constructed a bovine yeast artificial chromosome (YAC) library to provide a common resource for bovine genome research. We used leukocytes of a Japanese black bull (Bos taurus) as the DNA source, AB1380 for the yeast host, and pYAC4 for the vector. The library consists of 24,576 clones arranged in 256 96-well microtiter plates. An average insert size estimated from the analysis of 251 randomly selected clones was 480 kb. The rate of chimeric YACs evaluated by fluorescence in situ hybridization (FISH) analysis of 44 randomly selected clones was 36.4%. To estimate the number of genome equivalents, PCR-based screening was performed with 48 primer pairs and isolated 3.2 clones on average. In order to provide broad access for the scientific community, this library has been incorporated into the Reference Library system which provides high density filters for colony hybridization screening and a common database of the library.

摘要

我们构建了一个牛酵母人工染色体（YAC）文库，为牛基因组研究提供一个通用资源。我们使用日本黑牛（Bos taurus）的白细胞作为DNA来源，AB1380作为酵母宿主，pYAC4作为载体。该文库由24,576个克隆组成，排列在256个96孔微量滴定板中。通过对251个随机选择的克隆进行分析估计，平均插入片段大小为480 kb。通过对44个随机选择的克隆进行荧光原位杂交（FISH）分析评估，嵌合YACs的比例为36.4%。为了估计基因组当量数，使用48对引物进行基于PCR的筛选，平均分离出3.2个克隆。为了让科学界广泛获取该文库，此文库已被纳入参考文库系统，该系统提供用于菌落杂交筛选的高密度滤膜以及该文库的通用数据库。