西兰花端粒酶分离与活性测定探究

Isolation and Activity Evaluation of Broccoli Telomerase

植物端粒酶以自身的RNA亚基为模板,维持着染色体末端端粒的长度,从而使植物细胞可持续分裂。本研究以西兰花花蕾为材料,采用不同分子量、不同浓度的PEG分别提取端粒酶,根据PEG吸附蛋白质的原理,从具有端粒酶活性的组织中分离并富集端粒酶。结果显示:用PEG 6000提取西兰花端粒酶,且用100μg的酶量进行酶促反应时测得的端粒酶活性最高。以此为参数,经改进的TRAP法成功检测到:用0.2g PEG 6000/700μL洗脱液提取出的端粒酶活性很高。这些分离、检测技术为其它植物组织中端粒酶活性的检测和评价提供了新的途径,尤其对低活性的植物端粒酶检测具有重要意义,为深入研究端粒酶RNA的序列及结构奠定了基础。

Telomerase have its own RNA subunit as template for telomere extension, resulting in sta- bilizing the length of telomere at chromosomal end. Therefore, plant cells have the capacity to divide. This study uses the buds of the broccoli as materials, and adopts PEG of different molecular weight and differ- ent concentration to extract telomerase respectively. It realizes the purpose of separating and enriching te- lomerase from the tissue of telomerase activity according to the principle that PEG absorbs protein. The results show that the activity of the telomerase extracted by PEG 6000 is the highest when its content is 100μg for enzymatic reaction. Based on this, it detects the telomerase activity is the highest in the im- proved TRAP method when the eluent quantity amount is 0.2 g PEG6000/700 μL. These isolation and de- termination techniques could provide a new approach to determination and assessment of telomerase activity in other plant tissue. In particular, it has very important significance to determine the low activity of plant telomerase and provides basis for deeply researching sequences and structure of telomerase RNA.