春兰花发育时期的酵母双杂交文库的构建及评价

浙江农业学报

春兰花发育时期的酵母双杂交文库的构建及评价

（浙江省农业科学院园艺研究所，浙江杭州 310021）

出版日期:2015-03-25 发布日期:2015-03-31

Construction and evaluation of yeast two hybrid library of Cymbidium goeringii during flower development

（Institute of Horticulture， Zhejiang Academy of Agricultural Sciences， Hangzhou 310021， China）

Online:2015-03-25 Published:2015-03-31

摘要/Abstract

摘要： 为了研究春兰花发育相关蛋白之间的相互作用，文章使用gateway技术构建春兰花发育时期的酵母双杂交cDNA文库，文库质粒导入pGADT7\|Rec2\|DEST载体，并转化进酵母Y187中。检测表明，试验得到的初级文库库容量为147×107 cfu，酵母双杂交cDNA文库库容量高达254×107 cfu，文库的插入片段主要集中在1 000～2 000 bp。试验构建得到了高质量的春兰花发育时期酵母双杂交文库，为筛选相互作用的春兰花发育调控基因，研究春兰花发育的分子机制奠定了基础。

关键词: 春兰, 花发育, 酵母双杂交文库

Abstract: A yeast two hybrid cDNA library of Cymbidium goeringii during flower development was constructed using the CloneMinerTM cDNA Library Construction Kit. Purified plasmid DNA and linear vector （pGADT7\|Rec2\|DEST） were co\|transformed into yeast strain Y187. The results showed that the primary library contains 147×107 independent clones and the yeast two\|hybrid library contains 254×107 independent clones. The sizes of most inserts were 1\|2 kb in length. These results showed that the library was high quality， which is suitable for the screening of flower development related up\| and down\|regulated genes and provide an important foundation for the studies on the molecular mechanisms involved in flower development of Cymbidium goeringii.

Key words: Cymbidium goeringii, flower development, yeast two hybrid library

秦德辉，向林，李小白，吴超，孙崇波*. 春兰花发育时期的酵母双杂交文库的构建及评价 [J]. 浙江农业学报.

QIN De\|hui， XIANG Lin， LI Xiao\|bai， WU Chao， SUN Chong\|bo*. Construction and evaluation of yeast two hybrid library of Cymbidium goeringii during flower development [J]. .