Advantages and limitations of using Spm as a transposon tag.
Advantages and limitations of using Spm as a transposon tag.
复制标题
使用 Spm 作为转座子标签的优点和局限性。
DOI：
10.1007/978-1-4684-5550-2_11
复制
发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Burr,FA
中科院分区：
文献类型：
--
作者：
Cone,KC;Schmidt,RJ;Burr,B;Burr,FA
转座子标记已成为分离产物“低丰度”基因的首选方法。我们最近使用转座元件Spm来标记和克隆玉米调控位点。我们对Spm的选择是由几个因素决定的：在< 106种子的群体中，Spm的转位频率高到足以获得可检测的转位事件，进入影响籽粒性状的位点。尽管Spm在玉米基因组中的拷贝数很高，但通过使用甲基敏感限制性内切酶消化来自分离群体的dna，并与Spm特异性探针杂交，可以将插入到感兴趣基因中的Spm拷贝与其他Spm拷贝区分开来。由于迄今为止检测的所有Spm家族成员都具有DNA同源性，因此使用适当的探针进行杂交可以检测到自主和缺陷元件的插入。因此，如果一个可变的等位基因可以被证明是受Spm控制的，人们就可以有理由相信成功克隆了这个等位基因。
Transposon tagging has become the method of choice for isolating genes whose products are" in low abundance. We have recently used the transposable element Spm to tag and clone maize regulatory loci. Our choice of Spm was dictated by several factors: The frequency of transposition ofSpm is high enough to obtain detectable transposition events, into loci affecting kernel traits, in populations of< 106 seed. Although the copy number of Spm is high in the maize genome, insertions into the gene of interest can be distinguished from other Spm copies by digesting DNAs from segregating populations with methyl-sensitive restriction enzymes, and hybridizing with Spm-specific probes. Since all members of the Spm family thus far examined share DNA homology, hybridization with appropriate probes allows detection of insertions of both autonomous and defective elements. Thus, if a mutable allele can be shown to be under Spm control, one can be reasonably confident of successfully cloning that allele.