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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, in order to analyze the sequence and homology of the mat K gene collected from the west cactus in Haikou City, Hainan, total RNA was extracted from the cactus leaf using RNA plant Reagent kit and reverse transcribed into the first strand of c DNA. At the same time, a pair of conservative primers of the mat K gene of cactus was designed and the K gene of cactus mature enzyme was amplified by RT-PCR. The obtained sequences were compared by Blastn and found that 22 kernal genes The phylogenetic tree was constructed by using Blastn’s Distance tree of results. The results showed that the mat K gene of Hainan cactus has the highest homology with other mat C genes of 30 cacti reported, with homology above 99% Group, while the lowest homology with Opuntia basilaris (Gen Bank accession No.JF786750.1) was 96.6%. In this study, the gene sequence of mature cactus K gene was obtained by RT-PCR and the phylogenetic tree was constructed to confirm the homology of K gene of mature cactus. This result will provide scientific clues for the evolutionary research of Hainan cactus.