Joukhadar Reem, Jighly Abdulqader
University of Aleppo, Aleppo, Syria.
BMC Res Notes. 2012 Oct 5;5:556. doi: 10.1186/1756-0500-5-556.
Microsatellites, or simple sequence repeats (SSRs), are DNA sequences that include tandem copies of specific sequences no longer than six bases. SSRs are ubiquitous in all genomes and highly mutable.
Results from previous studies suggest that flanking regions of SSR are exhibit high stability in a wide range of organisms. We hypothesized that the SSRs ability to discard weak DNA polymerases could be responsible for this unusual stability. . When the weak polymerases are being decayed over SSRs, the flanking sequences would have higher opportunity to be replicated by more stable DNA polymerases. We present evidence of the molecular basis of our hypothesis.
The hypothesis could be tested by examining the activity of DNA polymerase during and after a number of PCRs. The PCR reactions should be run with the same SSR locus possessing differences in the SSR length. The hypothesis could also be tested by comparing the mutational rate of a transferred gene between two transformations. The first one has a naked T-DNA (transferred DNA), while the second one has the same T-DNA flanked with two SSRs.
In any transformation experiment, flanking the T-DNA fragment with SSR sequences would result in more stably transferred genes. This process would decrease the unpredictable risks that may occur because of the mutational pressure on this foreign segment.
微卫星,即简单序列重复(SSRs),是包含不超过六个碱基的特定序列串联重复的DNA序列。微卫星在所有基因组中普遍存在且高度可变。
先前研究的结果表明,微卫星的侧翼区域在广泛的生物体中表现出高度稳定性。我们假设微卫星丢弃弱DNA聚合酶的能力可能是造成这种异常稳定性的原因。当弱聚合酶在微卫星上衰减时,侧翼序列将有更高的机会被更稳定的DNA聚合酶复制。我们提供了支持我们假设的分子基础的证据。
可以通过在多次聚合酶链式反应(PCR)期间及之后检测DNA聚合酶的活性来检验该假设。PCR反应应使用具有不同微卫星长度差异的相同微卫星位点进行。该假设也可以通过比较两次转化之间转移基因的突变率来检验。第一次转化有一个裸露的T-DNA(转移DNA),而第二次转化有相同的T-DNA,两侧带有两个微卫星。
在任何转化实验中,用微卫星序列侧翼T-DNA片段将导致基因转移更稳定。这个过程将降低由于对该外源片段的突变压力可能出现的不可预测风险。