用PCR方法扩增深红红螺菌的draT基因

Amplification of draT Gene of Rhodospirillum rubrum by PGR Method

聚合酶链反应是最近几年分子生物学领域中一项重大的技术突破。典型的PCR引物内应含50％G+C,且没有自身互补序列,特别是在其3’-末端不应有内部二级结构。引物与靶DNA的退火温度一般为50℃,但当已有的引物不太符合要求时,就要调整退火温度。本文报道了用PCR方法扩增深红红螺菌draT基因,介绍了当引物GC含量较低时,可采用适当降低退火温度,然后梯度升温的方法获得PCR产物。

draT gene of Rhodospirillurn rubrum was amplified by PCR with different primers. When G+C content of the primer is lower,the products of PCR can be obtained by changing the annealing temperature from 50 ℃ for 2 munites to 37℃, 45℃ , 50℃ for 40 seconds each. The results of Dot-blot hybridization showed that the product of PCR is draT gene.