Isolation and characterization of SSR sequences from the genome and TAC clones of common wheat using the PCR technique.
We have developed the 2-step PCR method, a kind of suppression PCR procedure, to isolate simple sequence repeats (SSRs) from common wheat (Triticum aestivum L.) in a more convenient manner. This system requires neither genomic library screening nor the SSR-enrichment procedure. As a result, we designed 131 primer pairs based on isolated SSRs from not only genomic DNA, but also transformation-competent artificial chromosome (TAC) clones. It has been demonstrated that 34 of the 131 SSR markers developed were polymorphic among 8 wheat lines. Four of 34 polymorphic SSR markers were derived from TAC clones, indicating that this method could be applied to the targeted development of unique SSR markers in large genomic DNA libraries such as those composed of bacterial artificial chromosomes (BACs). A considerable number of isolated SSR clones had similarities with part of several long terminal repeats of retrotransposons (LTR-RTs) identified in various Triticeae genome sequences. Most of those SSRs showed smear amplification profiles, suggesting that a considerable number of dysfunctional SSRs originating from repetitive DNA components, especially LTR-RTs, might exist in the common wheat genome.
Koike
Michiya
M
Hokkaido Green-Bio Institute, Naganuma, Japan.
Kawaura
Kanako
K
Ogihara
Yasunari
Y
Torada
Atsushi
A
D001483:Base Sequence; D022201:Chromosomes, Artificial / Q000378:metabolism*; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D018744:DNA, Plant / Q000302:isolation & purification*; D005819:Genetic Markers; D018745:Genome, Plant; D018598:Minisatellite Repeats; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D014908:Triticum / Q000235:genetics*
D004251:DNA Transposable Elements; D018744:DNA, Plant; D005819:Genetic Markers