CAJ | 학술논문

为进一步探讨单胚RT-PCR技术的适用性,我们测试了3个特定的基因STM、MP和ASKη在拟南芥(Arabidopsisthaliana(L.)Heynh.)早期胚胎发生过程中的动态表达情况.进一步证实以该技术研究某些特定基因的表达动态是可行的.着重探讨了扩增产量与模板量的对应关系、基因组DNA的干扰以及DNase Ⅰ处理过程中的mRNA降解等主要技术问题.结果表明:材料制备时显微操作技术的稳定是确保单胚RT-PCR结果稳定的关键.应尽量缩短DNaseⅠ处理时间,以避免mRNA降解.
위진일보탐토단배RT-PCR기술적괄용성,아문측시료3개특정적기인STM、MP화ASKη재의남개(Arabidopsisthaliana(L.)Heynh.)조기배태발생과정중적동태표체정황.진일보증실이해기술연구모사특정기인적표체동태시가행적.착중탐토료확증산량여모판량적대응관계、기인조DNA적간우이급DNase Ⅰ처리과정중적mRNA강해등주요기술문제.결과표명:재료제비시현미조작기술적은정시학보단배RT-PCR결과은정적관건.응진량축단DNaseⅠ처리시간,이피면mRNA강해.
An extended investigation was carried out for further demonstrating single-embryo RT-PCR technique, which was previously established in our laboratory as a useful tool in the study of gene expression of Arabidopsis thaliana (L.) Heynh. Based on the previous work some important technical aspects in the application of the technique were mainly discussed in the present report. The dynamic expression of three selected genes, shoot-meristemless (STM), monppteros (MP) and Arabidopsis shaggy-like kinase etha (ASKη), was carefully compared during the embryogenesis. It was revealed that the technique was especially useful for tracing the expression dynamics of specific genes during embryogenesis. However,the quantity of starting material and the duration of DNase Ⅰ treatment are critical points for successful application of the technique. The best condition for eliminating the possible influence of genomic DNA on data analysis was also tested and discussed.