八棱海棠肌动蛋白基因(MrACT)的克隆及分析  

Cloning and analysis of MrACT from Malus micromalus

作　　者：徐锦涛[1] 章镇[1] 彭日荷[2] 熊爱生[2] 刘金戈[2] 周军[1] 蔡斌[1] 高峰[2] 付晓燕[2] 姚泉洪[2] 

机构地区：[1]南京农业大学园艺学院,南京210095 [2]上海市农业科学院生物技术研究所,上海201106

出　　处：《果树学报》2008年第3期289-292,共4页Journal of Fruit Science

基　　金：863项目(2006AA10Z117;2006AA06Z358);上海市科委重大基础项目(06DZ19103);上海市科技重点攻关项目(沪农科攻字(2006)第2-1号);上海市优秀学科带头人计划(06XD14017)

摘　　要：以八棱海棠(Malus micromalus Makino)为材料,抽取RNA,反转录成cDNA。采用cDNA末端快速扩增(RACE)方法,5’-RACE获得长度1121bp的片段、3’-RACE获得883bp的片段,再通过设计引物扩增得到八棱海棠中一种肌动蛋白基因的cDNA的全长序列。该序列全长1134bp,推测其编码377个氨基酸,等电点为5.16,其编码的氨基酸与拟南芥(Arabidopsis thaliana)actin7编码的氨基酸只有2个氨基酸差别。通过对八棱海棠MrACT基因的表达分析,发现在不同组织中,该基因的表达水平没有明显差异,本文为进一步利用RT-PCR技术,以MrACT基因为内标,研究八棱海棠其他基因的丰度打好基础。A ctin is a ubiquitous component of the plant cytoskeleton and participates in a number of important subcellular processes. The microfilament cytoskeleton is involved in cell division plane localization, cell elongation and cell shape determination. The plant aetins are made up of 376 or 377 amino acids and they are highly conservative. The high conservativeness accords with the actins as function of cytoskeleton. High-quality RNA was extracted from Malus micromalus seedlings. Then the eDNA was obtained through reverse transcription kit. An actin gene was obtained by rapid amplification of eDNA ends （RACE） from M. micromalus seedlings. Sequences analysis showed that the actin gene is 1 134 bp, which encodes 377 amino acids. Sequence analysis showed there are only two different amino acids between MrA CT and A rabidopsis thaliana actin 7. The predicted isoelectiic point of the protein is 5.16. Through RT-PCR assays we detected that the expression of MrA CT had no obvious difference in five kinds of tissues. We suggest that M. micromalus actin could be used as internal standard to study expression of other genes in M. micromalus.

关 键 词：八棱海棠 肌动蛋白基因 RACE 内标 

分 类 号：S661.19[农业科学—果树学]