数字农科院2.0

Construction of gateway-compatible yeast two-hybrid vectors for high throughput analysis of protein interaction

文献类型： 外文期刊

作者： Tingheng Zhu ; Weixia Wang *; Hann-lin Wong ; Xiao Yang ; Kun Wang ; Zhifeng Cui *

作者机构：

关键词： Yeast two-hybrid;gateway cloning technology;protein interaction

期刊名称： African Journal of Biotechnology

ISSN： 1684-5315

年卷期： 2010 年 9 卷 9 期

页码：

收录情况： SCI

摘要： Yeast two-hybrid system combined with the gateway technology will greatly facilitate the cloning of interested DNA fragment into yeast two-hybrid vectors and therefore increase the efficiency of yeast two-hybrid analysis. In this study, we constructed a pair of Gateway-compatible yeast two-hybrid vectors pBTM116GW and pVP16GW by introducing the gateway cassette (attR1-Cmr-ccdB-attR2) into the multiple cloning sites (MCS) of the previously described vectors pBTM116SS and pVP16S1, respectively. The applicability of newly generated vectors was tested by assaying the interaction between the kinase domain XA21K of rice (Oryza sativa) receptor like kinase XA21 and its interactor XB3. Since both Xa21K and Xb3 were cloned into a Gateway entry vector and then subcloned into pBTM116GW and pVP16GW by in vitro recombination with high efficiency, respectively, it demonstrated that the newly constructed gateway-compatible two-hybrid vectors will be useful in analysis of protein interactions in a high throughput way.

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