中国卤虫早期胚胎发育相关基因的研究——Orthodenticle基因克隆中简并引物的设计和优化

A study on the related genes of early embryonic development of Artemia sinica——Degenerate PCR primer design and improvement for Artemia sinica Orthodenticle clone

通过使用blastx、Blockmaker、CODEHOP和Primer Premier 5.0等在线网络工具和生物软件,针对中国卤虫Orthodenticle基因高度保守区域设计了简并引物.用所设计的简并引物克隆了中国卤虫Orthodenticle基因片段.在GenBank中以blastx方法进行比对,发现此段基因片段与美国卤虫Orthodenticle基因有高度的相似性.通过实验进一步证明,此种设计简并引物的方法可信性强,特异性高,能够快速得到满意的实验结果.

The Orthodenticle gene of Artemia sinica was cloned by using degenerate PCR. According to Orthodenticle gene highly conservative region, the degenerate primers was designed by using some network tools and biological softwares such as blastx, Blockmaker, CODEHOP and Primer Premier 5.0. Similarity comparison showed that the product of the cloned gene was very similar to EST of Orthodenticle gene of A. franciscana. It is considered that the degenerate primers obtained from this process design are practicable, and also get satisfied results and high positive rates.