摘要
分离克隆了龙眼(Dimocarpus longan Lour.)胚性愈伤组织Leafy Cotyledon 1(LEC1)基因,并分析该基因在龙眼体细胞胚胎(简称体胚)发生过程中的表达情况.采用RT-PCR结合RACE法及TAIL-PCR法,获得龙眼胚性愈伤组织LEC1基因的cDNA全长序列,运用生物信息学方法对该序列进行分析,并通过实时荧光定量PCR法(qRT-PCR)研究该基因在龙眼体胚发生过程中的表达.克隆得到LEC1基因803 bp的cDNA全长序列(GenBank检索号为GU584089.2),该cDNA的开放阅读框推定的氨基酸序列(含222个氨基酸)与其他植物LEC1具有较高同源性;该基因在龙眼体胚各阶段均有表达,在心形胚及鱼雷形胚阶段呈现高表达,整个变化趋势呈单峰型.
LEC1 gene was cloned from embryogenic callus and its expression patterns were determined during somatic embryogenesis in longan（Dimocarpus longan Lour.）.The RT-PCR（reverse transcription polymerase chain reaction）,RACE（rapid amplification of cDNA ends） and TAIL-PCR were used to isolate the complete cDNA sequence of LEC1 from longan embryogenic callus,and the bioinformatics methods were used to analyze the cDNA sequence and putative amino acid sequence.The full length LEC1 cDNA,which had been submitted to the DDBJ/EMBL/GenBank database（the accession number was GU584089.2）,was 803 bp,in addition,the putative protein had 222 amino acids,and the identity to the other polypeptides varied between 35%-70%.Finally,the qRT-PCR（real-time quantitative transcription PCR） method was used to determine the mRNA transcription levels of the gene.LEC1 was expressed at the different stages during somatic embryogenesis,showing a single peak curve with the relative higher mRNA transcription level of LEC1 gene at the heart embryo stage and the torpedo embryo stage.
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2011年第5期494-500,共7页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
福建省重大科技平台建设(2008N2001)
国家自然科学基金(31011787
30471204)资助项目
关键词
龙眼
体胚发生
LEC1
基因克隆
序列分析
实时荧光定量PCR
Dimocarpus longan
somatic embryogenesis
Leafy Cotyledon 1
gene cloning
sequence analysis
real-time quantitative transcription PCR
