Isolation and Characterization of New Maize B Sequences from a Microdissected Library
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Abstract
Understanding the evolution of the maize B chromosome requires insight into the molecular organization of a large number of B clones, which can be potentially obtained by microdissection of the chromosome. Yet, the microdissection protocols currently available are ineffective for a large-scale isolation. In an attempt to improve its efficiency, a protocol was adopted to screen a microdissected B library with probes prepared from the degenerate oligonucleotide primed-PCR product of genomic DNA. This protocol resulted in 59 new B clones, most of which were highly repetitive sequences located in various B regions but mostly in the heterochromatic blocks of the long arm. They also appeared in A chromosomes. Twenty-four of these were retrotransposons, ten knob, 18 noncoding sequences, and seven unknown sequences. The implication of the new B sequences on the B evolution is discussed.
Keywords
B chromosome DOP-PCR Evolution Microdissection Zea maysNotes
Acknowledgments
The authors would like to thank Dr. W.-H. Hsu and the Ministry of Education, Taiwan, R. O. C. under the ATU plan for financial support on part of this study and H.-H. Yang for technical assistance.
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