猪染色体的显微切割：用于构建染色体涂染探针和文库的整条染色体、染色体臂及条带的切割

Microdissection of pig chromosomes: dissection of whole chromosomes, arms and bands for construction of paints and libraries.

作者信息

Chaudhary R, Kijas J, Raudsepp T, Guan X Y, Zhang H, Chowdhary B P

机构信息

Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Uppsala, Sweden.

出版信息

Hereditas. 1998;128(3):265-71. doi: 10.1111/j.1601-5223.1998.00265.x.

DOI:10.1111/j.1601-5223.1998.00265.x

PMID:9760875

Abstract

Chromosome microdissection is an important means to efficiently generate a large number of markers from a desired region of a genome. The present study was designed to initiate microdissection and amplification of DNA from whole chromosomes, arms, or bands of porcine chromosomes. The following pig (SSC) chromosomes/segments were scraped: SSC1p, SSC1q26-q2.13, SSC2q11-q14, SSC4q12-q25, SSC13, SSC13q12-q31, SSC13q32-q43, SSC13q32-q43, SSC15, and SSC16q21-q23. After amplification and PCR-labelling, the DNA from the dissected segments were painted back to normal metaphase chromosomes to test their identity. Microdissection of some of the segments (on SSC4, 13 and 15) coincides with the mapping of economically important traits. As a first step towards generation of markers, microcloning of amplified product from SSC1p and SSC15 was carried out. The libraries were screened with a (GT)15 oligonucleotide probe. Future prospects of such a work in farm animals are discussed.

摘要

染色体显微切割是从基因组的特定区域高效生成大量标记的重要手段。本研究旨在启动对猪染色体的整条染色体、染色体臂或染色体带的显微切割及DNA扩增。刮取了以下猪（SSC）染色体/片段：SSC1p、SSC1q26 - q2.13、SSC2q11 - q14、SSC4q12 - q25、SSC13、SSC13q12 - q31、SSC13q32 - q43、SSC13q32 - q43、SSC15以及SSC16q21 - q23。扩增及PCR标记后，将切割片段的DNA回涂到正常中期染色体上以检测其同一性。部分片段（SSC4、13和15上的）的显微切割与经济重要性状的定位相吻合。作为生成标记的第一步，对SSC1p和SSC15的扩增产物进行了微克隆。用（GT）15寡核苷酸探针筛选文库。讨论了此类工作在农场动物中的未来前景。