5种植物花瓣ACS基因的克隆与分析

ACS gene cloning and analysis from petals in five plants

分析已发表的植物1-氨基环丙烷-1-羧酸(ACC)合成酶(ACS)基因序列,设计一对简并引物,在随机选择的非洲菊、月季、桂花、茶梅和山茶等5种植物花瓣cDNA中进行PCR扩增.结果表明,在5种植物花瓣中都能扩增出约800 bp的特异片段,登录GenBank发现均未被注册,因此将序列提交到GenBank并获得了序列号.在NCBI上进行Blast比对,发现所克隆的5个ACS基因均与其他植物花瓣的ACS基因有一定的序列相似性,最高达94%,最低为80%.将5个ACS基因的片段进行多序列比较分析,发现茶梅ACS基因与山茶ACS基因的序列差异最小,序列相似性达99%.本试验设计的ACS基因简并引物在植物中有一定的通用性,后续可以适当调整引物的简并度,以得到高通用性的简并引物.

Based on the published sequences of plant ACS genes, a pair of degenerate primers was designed to clone ACS genes from petals in five plants. The results indicated that specific fragments of about 800 hp could be amplified from all the five plants and the sequences were not registered on GenBank database before this work. And then the sequences were submitted to GenBank database and the accessing numbers were offered. The five sequences show higher identities through NCBI Blast anal）sis, and the highest and lowest percentage are respectively 94% and 80%. It was found that the sequence difference between Camellia sasanqua ACS gene and C.japomica ACS gene was the smallest by multiple sequence alignments and the identity is 99%. The designed degenerate prim- ers in the study have some generality in plants, and the primer of degeneracy would be adjusted in future to obtain higher unlversalizable degenerate primers.