%0 Journal Article
%T Microdissection of chromosome 7B of common wheat and cloning of low-copy specific DNA sequences
%A Bao Liu
%A Junkang Rong
%A Yingshan Dong
%A Fangpu Han
%A Zhenlan Liu
%A Mengyuan He
%A Baiqu Huang
%A Shui Hao
%J Chinese Science Bulletin
%@ 1861-9541
%D 1999
%I 
%R 10.1007/BF03182725
%X The 7B chromosome of common wheat was microdissected from pollen mother cells of the 78 monosomic line of common wheat cv. Chinese Spring (CS). After proteinase K and DNA topoisomerase I treatments, the isolated chromosomes were subjected to 1¨C3 rounds of DOP-PCR amplification, which produced continuous DNA fragments ranging from 150 to 700 bp. Genomic Southern hybridization confirmed that the PCR products were originated from the wheat genome. Cloning of portion (>200 bp) of the 3rd round DOP-PCR products (50 ¦ĚL could generate about 20 000 recombinant clones. Characterization of 50 randomly chosen clones indicated that 21 clones produced discrete PCR products with the sue of 240¨C600 bp. Dot-blot hybridization showed that among the 21 clones, 11 (ˇ«55%) were of low-copy nature while 10 (ˇ«45%) were repetitive. Southern hybridization with the complete set of the CS ˇ°nullisomic-tetrasomic (NT)ˇ± lines demonstrated that all the 6 low-copy clones were specific to either chromosome 78 or the 7th homoeologous group, whereas the 3 arbitrarily chosen repetitive clones were non-specific, disperse sequences.
%K wheat
%K chromosome microdissection
%K DOP-PCR
%K specific DNA sequences
%U http://link.springer.com/article/10.1007/BF03182725