Cole K P, Blakeley S D, Dennis D T
Department of Biology, Queens University, Kingston, Ontario, Canada.
Gene. 1992 Dec 15;122(2):255-61. doi: 10.1016/0378-1119(92)90213-9.
The polymerase chain reaction (PCR) has been used to generate a series of overlapping genomic clones representing 43 bp of 5' untranslated sequence, 63 bp of 3' untranslated sequence and the entire coding sequence of the gene encoding potato cytosolic pyruvate kinase (PKc). This portion of the gene is approximately 4.5 kb in length and is interrupted by three introns, one of which is present in the 5' untranslated region. Southern blot analysis indicates that PKc is encoded by a small gene family, and sequence data from a number of PCR-derived genomic clones indicate that there are as many as six PKc genes. Sequence differences between the PCR-generated genomic clones and a PKc cDNA clone are discussed with respect to the fidelity of Taq polymerase. An alignment of intron placement in the potato PKc gene with intron placement in PK genes from other sources indicates that two of the potato introns correspond to intron positions in other species.
聚合酶链反应(PCR)已被用于生成一系列重叠的基因组克隆,这些克隆代表了43个碱基对的5'非翻译序列、63个碱基对的3'非翻译序列以及编码马铃薯胞质丙酮酸激酶(PKc)的基因的完整编码序列。该基因的这一部分长度约为4.5千碱基对,被三个内含子打断,其中一个内含子存在于5'非翻译区。Southern印迹分析表明PKc由一个小基因家族编码,来自多个PCR衍生基因组克隆的序列数据表明多达有六个PKc基因。就Taq聚合酶的保真度而言,讨论了PCR生成的基因组克隆与PKc cDNA克隆之间的序列差异。马铃薯PKc基因中内含子的位置与其他来源的PK基因中内含子的位置比对表明,马铃薯的两个内含子与其他物种的内含子位置相对应。