大麦1H特异性CAPs标记和ASA标记的创制

选取大麦1H染色体的STS标记MWG913特异性扩增小麦,把得到的片段进行克隆.用Taq酶切分类并测序,把得到的序列同大麦的序列进行比较.依据比较结果,选取对大麦特异的内切酶,用该酶来酶切大麦、小麦、黑麦、长穗偃麦草、中间偃麦草、簇毛麦的MWG913扩增产物,获得对大麦1H染色体特异的CAPs标记.同时,依据酶切位点碱基的差异设计引物对扩增的产物进行第二次扩增,得到该位点的一对染色体特异性ASA标记.

The Creation of 1H Chromosome-specific CAPs and ASA Markers of Barley

Sequence-Tagged Site(STS) marker is a kind of important marker for identifying heterogeneous chromosome on account of its simplicity, speediness and specificity. There are many STS markers in use, which changed from the RFLP and RAPD markers. But some of those from RFLP markers cannot directly be used for identifying the specific chromosomes, such as MWG913. Amplified wheat genome by MWG913, a STS marker of barley's 1H chromosome, then cloned the products. These clones were classified by restricted enzyme Taq I and then sequenced. On the basis of the results of these sequences compared with barley's (obtained from the GenBank)., two restricted enzyme specifying on 1H chromosome, EcoR I and Pvu I, were chosen to digest the PCR products of T aestivum, Secale cereale, Hordeum vulgare, Agropyron intermedium, Haynaldia villosa, Thinopyrum elongatum and identified their specilization to Hordeum vulgare genome. Furthermore, according to the difference of Pvu I recognition site, the second PCR primers were designed and identified its specificity to Hordeum vulgare genome by PCR Thus 1H chromosomespecific CAPs and ASA markers of barley were obtained.