摘要
单染色体分离、扩增和克隆技术对植物的遗传研究（尤其对那些遗传研究基础薄弱的植物而言）具有重要的应用价值。本研究用玻璃针显微分离法成功分离出长果种黄麻（Corchorus olitoriusL．）品种闽引一号的单染色体80条。用LAM-PCR或DOP-PCR对单染色体DNA进行两轮扩增，所包含的DNA片段长度分别为250-2000bp和100-600bp。共建立了50个单染色体未克隆文库。Southern杂交表明，获得的扩增片段确实来自黄麻基因组。对单染色体DNA文库应用的有关问题进行了讨论。
The technology of isolation, amplification and cloning of single chromosome is of significant value in plant genetic research, especially for those plants with poor genetic research basis. However, the technology has not been widely applied to plants because most plants have small chromosomes that are difficult to be distinguished. To investigate the feasibility and possibility of applying the technology to plants with small chromosomes, we experimented on the construction of uncloned single chromosomal DNA libraries using jute as a model, which has typical small chromosomes. We successfully isolated 80 single chromosomes from cultivar Minyinl of jute （ Corchours olitorius L. ） by micromanipulation with glass needles and amplified individual chromosomes by two-round LAM-PCR or DOP-PCR, from which DNA fragments ranging 250 - 2 000 bp or 100 - 600 bp were obtained. Southern hybridization with a-32p-dCTP labeled genomic DNA confirmed that the PCR products were really from the jute genome. Hence, we acquired uncloned DNA libraries of single chromosomes of jute. A total of 50 uncloned single-chromosome DNA libraries were established. Our research suggests that isolation individual small chromosomes with glass needles and amplification of DNA fragments from them with LAM-PCR or DOP-PCR for constructing uncloned single chromosome DNA libraries in plants is feasible. Issues concerning the application of uncloned single-chromosome DNA libraries are discussed.
出处
《作物学报》
CAS
CSCD
北大核心
2006年第8期1184-1187,共4页
Acta Agronomica Sinica
关键词
黄麻
单染色体
分离
未克隆文库
Jute
Single chromosome
Isolation
Uncloned library
