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Isolation of the genes encording DNA binding protein interacting with 5^ upstream region of the rice waxy gene by using yeast one-hybrid system

Chen Li, Xing Yanyan, Zhang Jingliu, Wang Zongyang, Hong Mengmin

Shanghai Institute of Plant Physiology,Chinese Academy of Sciences.Shanghai 200032

Abstract

A 31 bp DNA fragment within the 5^ upstream region of rice waxy gene was used as a bait in a yeast one-hybrid system to isolate the genes encoding trans-acting factors regulating rice waxy gene expression. This yeast one-hybrid system consists of two plasmids: the p178 plasmid consisted of two copies of 31 bp oligonucleotids adjacent to the mini-promoter Pcyc1 directing lacZ gene expression, in the second pPC86 plasmid, a rice cDNA library was ligated to a gene encoding GAL4 transactivating domain as a translational fusion to form a rice cDNA-GAL4 library. A yeast strain harboured the p178 plasmid was then transformed with the pPC86 plasmid containing cDNA-GAL4 library, and grown on medium containing X-gal to select blue colonies. After several round selections, thirteen positive clones were obtained. These thirteen cDNA clones can be divided into four groups according to their homology by Southern blot analysis.

Key words: Rice;Waxy gene;Binding protein;Yeast one-hybrid system

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Citing This Article：

Chen Li, Xing Yanyan, Zhang Jingliu, Wang Zongyang, Hong Mengmin. Isolation of the genes encording DNA binding protein interacting with 5^ upstream region of the rice waxy gene by using yeast one-hybrid system. Acta Physiol Sin 1999; 51 (2): (in Chinese with English abstract).