Čeština
EnglishAnalysis of nucleotide sequence of Pisum sativum chromosome-5-specific library
Wolko, L.; Vrána, Jan; Doležel, Jaroslav; Slomski, R.
In Proceedings of the International Workshop 'Understanding the Plant Genomeö. Poznan : Institute of Plant Genetics PAS : 79-88, 2004
Keywords: Pisum sativum; flow-sorted chromosomes; chromosome-5-specific DNA library
Abstract: Sequencing of clones derived from short insert chromosome-5-specific DNA library of pea may provide valuable information of chromosome structure. Our library was constructed by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) amplification using DNA of flow-sorted chromosomes as template. Analysis of nucleotide sequence of 200 randomly selected clones indicated that about 20% of clones were overrepresented in the library. The nucleotide sequence of 155 original chromosome-specific fragments was revealed. Sequence homology analysis showed that minority of fragments were similar only to uncharacterised ge-nomic sequences of different plant species. However, the set of sequences revealed different ranges of similarity to plant genes and different types of moderate repetitive elements. The results suggested equal reflection of real genome composition in the library clones, with the exception of short tandem repeats sequences. Shotgun characterisation implied a rather decreased content of microsatellite repeats in the library. Representative character of clones makes the library a valuable tool for draft large genome survey that can be useful as a source of chromosome specific markers.
DOI:
IEB authors: Jaroslav Doležel
In Proceedings of the International Workshop 'Understanding the Plant Genomeö. Poznan : Institute of Plant Genetics PAS : 79-88, 2004
Keywords: Pisum sativum; flow-sorted chromosomes; chromosome-5-specific DNA library
Abstract: Sequencing of clones derived from short insert chromosome-5-specific DNA library of pea may provide valuable information of chromosome structure. Our library was constructed by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) amplification using DNA of flow-sorted chromosomes as template. Analysis of nucleotide sequence of 200 randomly selected clones indicated that about 20% of clones were overrepresented in the library. The nucleotide sequence of 155 original chromosome-specific fragments was revealed. Sequence homology analysis showed that minority of fragments were similar only to uncharacterised ge-nomic sequences of different plant species. However, the set of sequences revealed different ranges of similarity to plant genes and different types of moderate repetitive elements. The results suggested equal reflection of real genome composition in the library clones, with the exception of short tandem repeats sequences. Shotgun characterisation implied a rather decreased content of microsatellite repeats in the library. Representative character of clones makes the library a valuable tool for draft large genome survey that can be useful as a source of chromosome specific markers.
DOI: