海南仙人掌matK基因的克隆和序列分析

Cloning and Sequence Analysis of matK Gene from Cactus

2015年12月,为了分析采自海南海口城西仙人掌mat K基因的序列及同源关系,利用RNA plant Reagent试剂盒提取仙人掌叶总RNA,反转录成第一链c DNA。同时设计仙人掌成熟酶K基因(mat K)的1对保守引物,利用RT-PCR的方法扩增仙人掌成熟酶K基因,所得序列经Blastn进行比对,发现与已报道的22个仙人掌mat K基因核苷酸序列一致;利用Blastn的Distance tree of results构建系统进化树,结果显示海南仙人掌mat K基因与其它报道的30个仙人掌mat K基因同源性最高,同源性在99%以上,属于同一组,而与Opuntia basilaris(Gen Bank accession No.JF786750.1)同源性最低,为96.6%。本研究利用RT-PCR方法获得了仙人掌成熟酶K基因序列,同时构建的系统进化树确定了该仙人掌成熟酶K基因的同源关系。本结果将为海南仙人掌的进化研究提供科学线索。

In December 2015, cactus leaf from Chengxi, Haikou, Hainan. was collected. In order to analyze the homology between Chengxi cactus and other cactuses, the total RNA was extracted using RNA plant Reagent kit and reverse transcribed into the first strand cDNA. Conserved pair primers of marK gene were designed, and used for obtaining matK gene fragment by RT-PCR amplification method. The Blastn of nucleotide sequences showed that matK gene （Chengxi cactus） was identical to 22 marK genes from GenBank. The distance tree of results program on blastn website was used to constructed phylogenetic tree. The results showed that marK gene （Chengxi cactus） was at least 99% similar to 30 other reported mark genes, belonging to the same group; while mark gene （Chengxi cactus） was 96.6% similar to Opuntia basilaris （GenBank accession No. JF786750.1）. In this study, marK gene of cactus was obtained by RT-PCR method, and the relationship between Chengxi cactus and other cactuses was also determined. The results would provide a clue to the evolution of cactus in Hainan.