Inheritance and molecular variations of PCR-SSCP fragments in pedunculate oak (Quercus robur L)
Inheritance and molecular variations of PCR-SSCP fragments in pedunculate oak (Quercus robur L)
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DOI：
10.1007/bf00223175
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发表时间：
1996-08-01
影响因子：
5.4
通讯作者：
Kermer, A
中科院分区：
文献类型：
--
作者：
Bodenes, C;Laigret, F;Kermer, A
对栓皮栎(Quercus I robur L.)3个全同胞家系的6个扩增片段(250~800bp)的单链构象多态性(SSCP)图谱进行了分析。还有他们的父母。在这6个片段中，4个片段是多态的，1个片段表现出复杂的图谱，这些图谱不会因SSCP条件的变化而改变。分析片段的条带数量在个体之间变化在2到4条之间，与片段大小无关。分离数据表明，树间带数的差异只能归因于等位基因组成(纯合子与杂合子)：表现两条带的基因型推定为纯合子，而表现三条或四条带的基因型则为杂合子。在每个多态片段的所有杂交组合中都观察到孟德尔比例。在一个杂交中，我们可以清楚地识别一个零等位基因，这是由于一个引物点可能发生了突变。单碱基突变和短插入缺失是不同等位基因间SSCP多态的分子原因。在基因多样性研究或基因定位中，建议使用SSCP作为一种技术来识别PCR片段的共显性标记(最高可达800bp)。
Single-strand conformaiton polymorphism (SSCP) profiles of six PCR-amplified fragments (250-800 bp) were analyzed in three full-sib families of pedunculate oak (Quercus I robur L.) and their parents. Among the six fragments, four were polymorphic and one exhibited complex patterns that were not changed by varying the SSCP conditions. The number of bands for the analyzed fragments varied between two and four among individuals regardless of fragment size. As shown by segregation data, the variation in the number of bands between trees could only be attributed to the allelic composition (homozygotes vs heterozygotes): a genotype that exhibited two bands was presumptively homozygous, wheras a genotype exhibiting three or four bands was heterozygous. Mendelian proportions were observed in all crosses for each polymorphic fragment. In one cross, we could clearly identify a null allele due to a possible mutation at a primer site. Single-base mutations and short insertion-deletions were shown to be the molecular causes of the SSCP polymorphism observed between different alleles. The use of SSCP as a technique to identify co-dominant markers of PCR fragments (up to 800 bp) is recommended for gene diversity studies or for gene mapping.