紫草LeEXT-1基因的克隆、序列及表达分析  

Cloning,sequence and expression analysis of LeEXT-1 gene in Lithospermum erythrorhizon

作　　者：汤程贻[1] 殷晶晶[1] 邹爱兰[1] 戚金亮[1] 杨永华[1] 

机构地区：[1]南京大学生命科学学院/医药生物技术国家重点实验室,江苏南京210093

出　　处：《南京农业大学学报》2013年第1期13-18,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31171161;31170275;31071082);教育部创新团队项目(IRT1020);江苏省自然科学基金项目(BK2010053;BK2011414)

摘　　要：以硬紫草细胞为材料,采用快速扩增cDNA末端法(rapid amplification of cDNA ends,RACE)克隆了由抑制消减杂交(suppression subtractive hybridization,SSH)和cDNA芯片技术筛选获得的在黑暗条件下优先表达的LeEXT-1基因的全长cD-NA。生物信息学分析结果表明,该cDNA全长为1 077 bp,编码1个含247个氨基酸的蛋白,与编码伸展蛋白(extensin)的基因具有高度的同源性。基因表达分析结果表明,当紫草细胞从B5培养基转到M9培养基时,LeEXT-1基因在2 h内被快速诱导表达,随后又逐渐降低到一个较低的水平。该基因的克隆为进一步研究伸展蛋白在紫草宁合成调控中的作用奠定了基础。In this study,the full-length cDNA of LeEXT-1 gene,which was identified by suppression subtractive hybridization（SSH）and cDNA microarray as a gene preferentially expressed in the dark,was cloned by using rapid amplification of cDNA ends（RACE）method from Lithospermum erythrorhizon cells.Bioinformatic analysis results showed that the full-length cDNA of LeEXT-1 was 1 077 bp,encoding a peptide of 247 amino acids with high similarity to extensin proteins.The expression analysis result showed that the mRNA level of LeEXT-1 was rapidly induced within 2 h when cells were transferred from B5 to M9 medium and then decreased to a low level in the following hours.The successful cloning of LeEXT-1 gene provides a foundation for understanding the function of extensin in regulating the biosynthesis of shikonin.

关 键 词：紫草 紫草宁 基因克隆 基因表达 伸展蛋白 

分 类 号：Q781[生物学—分子生物学]