青蒿鲨烯合酶cDNA的克隆与序列分析

cDNA Cloning and Sequencing of Squalene Synthase of Artemisa apiacea

[目的]对青蒿鲨烯合酶进行cDNA克隆,并进行序列分析。[方法]根据GenBank上已发表的鲨烯合酶cDNA基因序列设计1对特异性引物,提取青蒿细胞总RNA,运用RT-PCR扩增出鲨烯合酶基因。将其与pMD19-T载体连接,并对克隆片段序列进行分析。[结果]SS基因全长1257bp,编码418个氨基酸;同源性分析表明,青蒿SS基因序列与人参、积雪草、金铁锁、大豆、辣椒、百脉根、远志、玉米、褐家鼠、小鼠以及人相应序列的同源性分别为77.21%、76.11%、75.66%、74.62%、73.83%、74.46%、73.03%、64.39%、52.22%、50.51%和50.12%;氨基酸同源性分别为79.43%、79.00%、75.84%、79.43%、77.27%、77.27%、77.03%、66.03%、40.65%、40.19%和40.09%。[结论]青蒿鲨烯合酶cDNA的成功克隆,为进一步研究SS基因结构、基因表达与调控提供了重要依据。

[Objective] cDNA from squalene synthase was cloned and sequenced.[Method] A pair of specific primers were designed according to the cDNA gene sequence of squalene synthase published in GenBank.Total RNA was extracted from the cell of Artemisia apiacea.The genes of squalene synthase were amplified by using RT-PCR.It was connected with pMD19-T vector and the cloned fragment sequences were analyzed.[Result] SS gene with the whole length of 1257 bp was amplified and the fragment encoded 418 amino acids.The homology of SS gene between Artemisa apiacea and Panax ginseng,Centella asiatica,Psammosilene tunicoides,Glycine max,Capsicum annuum,Lotus japonicus,Polygala tenuifolia,Zea mays,Rattus norvegicus,Mus musculus and human were 77.21%,76.11%,75.66%,74.62%,73.83%,74.46%,73.03%,64.39%,52.22%,50.51% and 50.12%,respectively.The homology of the deduced amino acid were 79.43%,79.00%,75.84%,79.43%,77.27%,77.03%,66.03%,40.65%,40.19% and 40.09%,respectively.[Conclusion] cDNA of squalene synthase from Artemisia apiacea was successfully cloned,which provided important base for further study on the structure,gene expression and regulation of SS gene.