【摘要】 利用RT-PCR技术,首次克隆了斑茅1-氨基环丙烷-1-羧酸氧化酶基因序列.该片段长度为1123bp的片段,包括1个编码322个氨基酸的完整开放读码框,包括具有与异青霉素合成酶功能区类似的氧化功能PcbC区,和1个Fe2+和CO2活性中心。应用实时荧光PCR技术分析了1-氨基环丙烷-1-羧酸氧化酶基因在聚乙二醇胁迫下4个时段的表达。结果表明,聚乙二醇胁迫2h时,该基因在叶片明显上调表达,4h后的表达趋向平缓。本研究为1-氨基环丙烷-1-羧酸氧化酶基因进一步用于基因工程研究奠定基础,同时也初步揭示1-氨基环丙烷-1-羧酸氧化酶基因的表达与水分胁迫相互关系,为探讨乙烯对水分胁迫响应的分子机制提供了初步依据。更多还原

【Abstract】 l-aminocyclopropane-1-carboxylate oxidase gene(aco)was isolated from Erianthus arundinaceus by RT-PCR.Sequencing and BLASTX analysis in NCBI revealed that the cloned aco is 1 123 bp long and contains the ORF(open reading frame)of 966 bp which encodes 322 amino acids.Conserved domains were searched and three domains were found:one PcbC domain that possesses oxidation ability,and bivalent Fe2+ and active centre of CO2 in PcbC domain.Real-time fluorescent quantitative PCR analysis showed that the expression profile of aco under 4 stages of PEG stress was irregular at the early stage,and low at the consequent stages.At the stage of 2 hours of PEG stress,the expression was obviously higher,which was 5.4 times as that of the control,and since then became lower,but was still positively higher than that of the control,no matter at 4 hours or 12 hours of stresses.It is interesting to note that the expression of aco and control genes were almost the same with slight discrepancy.The present study confirmed the sensitivity of aco to external factors,and primarily its relationship with water stress.更多还原