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利用RecA蛋白的同源重组特性,以生物素标记的STK类抗性基因PCR扩增产物为探针,对小粒野生稻DNA文库进行富集,构建了库容量为1 710个的STK类抗性基因富集文库,通过菌落原位杂交筛选获得21个阳性克隆.其中10个阳性克隆进行了两端测序,生物信息学分析表明:有3个阳性克隆可分别定位于粳稻抗性基因附近和抗性基因内,可能是新的候选抗性基因;有2个阳性克隆可以在粳稻上定位,但周围无抗性基因片段;其它5个阳性克隆由于与栽培稻有多处匹配,不能精确定位。
Using the homologous recombination characteristic of RecA protein, the DNA library of small-sized wild rice was enriched with the PCR amplification product of biotin-labeled STK-resistant gene as a probe to construct a STK-type resistance library with a storage capacity of 1 710 Twenty-one positive clones were screened by colony in situ hybridization and 10 positive clones were sequenced at both ends. Bioinformatics analysis showed that three positive clones could be located in the vicinity of japonica resistance genes and resistance Two positive clones could be located in japonica rice without surrounding resistance gene fragments. The other five positive clones could not be precisely located due to many matches with cultivated rice.