北方农业学报2016，Vol.44Issue(5)：8-14,7.DOI:10.3969/j.issn.2096-1197.2016.05.02
拟南芥低钙诱导cDNA文库的构建
Construction of cDNA library induced by low Ca2+in Arabidopsis thaliana
摘要
Abstract
Calcium is an indispensable nutrient element for plant growth and development. The calcium ion (Ca2+) is a versatile intracellular messenger that is involved in numerous signalling pathways. In order to explore the regulatory mechanism of calcium homeostasis in plants, we isolated total RNA from Arabidopsis thaliana seedlings induced with low Ca2+. First, SMART cDNA synthesis technology was used to create a cDNA pool with end sequences homologous to the prey vector pGADT7-Rec. Then we co-transformed Y187 Yeast Strain and allowed the yeast to perform a recombination step between the linear prey vector and the cDNA. The cDNA library induced by low Ca 2+ in Arabidopsis thaliana was thus constructed. The library capacity is about 3 ×109 cfu/mL. The length of the insertion fragments ranges from 200 to 2 700 bp with an average length of 760 bp and the positive clone rate is 79.2%. These data suggest that the cDNA library is in high quality and suitable for screening the interacting proteins in Arabidopsis thaliana.关键词
拟南芥/Ca2+/cDNA文库Key words
Arabidopsis thaliana/Ca2+/cDNA library分类
农业科技引用本文复制引用
高天,铁英,王妍,祁智..拟南芥低钙诱导cDNA文库的构建[J].北方农业学报,2016,44(5):8-14,7.基金项目
国家自然科学基金项目（31171364）；内蒙古自治区2015年博士研究生科研创新项目 （）
