枇杷LEAFY同源基因的克隆及序列分析

Cloning and sequence analysis of LEAFY homologous gene from loquat

为从分子水平研究枇杷成花的机理,通过分析植物花分生组织决定基因LEAFY(LFY)同源基因的保守区序列,设计了1对简并引物,用PCR方法从枇杷栽培品种‘早钟6号’基因组DNA中扩增出1个1317bp的片段,把该片段克隆到pUCm T载体,测序和序列分析结果表明获得了枇杷LFY同源基因(ejLFY) 3′端的1个片段,该片段有1个911bp的内含子,编码区共编码130个氨基酸,其序列已经在GenBank中登记(登录号为AY5 5 1183) .在GenBank中进行同源性搜索,发现该基因片段与其他作物中已经报道的LFY同源基因的同源性大都在94 %以上,特别是与同属于蔷薇科的苹果的同源性最高,达到98%的同源性。

In order to study the molecular mechanism of floral formation on loquat, a pair of degenerate primers was designed according to the conservative regions of the plant floral meristem identity gene-LEAFY (LFY) homologous genes. A (1 317) bp fragment of LFY homolog gene was amplified by polymerase chain reaction (PCR) using the genomic DNA of 'Zao Zhong No.6' loquat cultivar. The fragment was cloned into pUCm-T vector, and then sequenced. The results suggest that a fragment in the 3′-end of LFY homologous gene named ejLFY was obtained. The sequence analysis indicated that there was an intron of 911 bp in the fragment, and the exons encoded 130 amino acids. The ejLFY gene was registered in GenBank with the accession number AY551183. After the deduced amino acid sequence of the fragment was submitted to GenBank to blast, it was found that the homology reached 94% to most of the other LFY homologous gene, especially to Malus family-apple, the homology reached the highest level (98%). This result suggested that ejLFY gene might have the same function as other LFY homologous gene.