ISOLATION OF MOLECULAR MARKERS FROM SPECIFIC CHROMOSOMAL INTERVALS USING DNA POOLS FROM EXISTING MAPPING POPULATIONS
ISOLATION OF MOLECULAR MARKERS FROM SPECIFIC CHROMOSOMAL INTERVALS USING DNA POOLS FROM EXISTING MAPPING POPULATIONS
复制标题
DOI：
10.1093/nar/19.23.6553
复制
发表时间：
1991-12-11
影响因子：
14.9
通讯作者：
TANKSLEY, SD
中科院分区：
文献类型：
--
作者：
GIOVANNONI, JJ;WING, RA;TANKSLEY, SD
我们提出了一个通用的方法，分离分子标记特定于任何区域的染色体使用现有的映射人口。 从现有作图群体的成员构建两个DNA库，所述DNA库来自由两个或更多个连锁RFLP标记限定的靶向染色体区间的相对等位基因纯合的个体。 然后使用随机寡核苷酸引物和PCR筛选DNA池的多态性（1）。 多态性DNA条带应代表所选区间内或附近的DNA序列。 我们使用两个基因组间隔在番茄中测试了这种方法，所述基因组间隔含有负责调节蒂脱落（无节）和果实成熟（非成熟）的基因。 用200个随机引物筛选每个区间含有7至14个F2个体的DNA池。 三个多态性标记，从而确定，其中两个随后被证明是紧密连锁的选定的间隔。 第三个标记定位在同一条染色体（11）上，但距离所选区间45 cM。 该方法的一个特别吸引人的属性是，单个作图群体可用于靶向基因组中的任何区间。 虽然这种方法已在番茄中得到证实，但它应该适用于任何有性繁殖的生物，其分离群体被用于构建遗传连锁图谱。
We present a general method for isolating molecular markers specific to any region of a chromosome using existing mapping populations. Two pools of DNA from individuals homozygous for opposing alleles for a targeted chromosomal interval, defined by two or more linked RFLP markers, are constructed from members of an existing mapping population. The DNA pools are then screened for polymorphism using random oligonuclotide primers and PCR (1). Polymorphic DNA bands should represent DNA sequences within or adjacent to the selected interval. We tested this method in tomato using two genomic intervals containing genes responsible for regulating pedicle abscission (jointless) and fruit ripening (non-ripening). DNA pools containing 7 to 14 F2 individuals for each interval were screened with 200 random primers. Three polymorphic markers were thus identified, two of which were subsequently shown to be tightly linked to the selected intervals. The third marker mapped to the same chromosome (11) but 45 cM away from the selected interval. A particularly attractive attribute of thie method is that a single mapping population can be used to target any interval in the genome. Although this method has been demonstrated in tomato, it should be applicable to any sexually reproducing organism for which segregating populations are being used to construct genetic linkage maps.