Cloning and Yeast Self-activated Detection Analysis of GhMADS43 Gene in Cotton

doi:10.7668/hbnxb.20191274

Abstract

Abstract: In order to elucidate the cotton GhMADS43 gene in shoot apical meristem and floral meristem development and to screen for proteins that interact with this geneby yeast two hybrid system.This study cloned the coding sequences of GhMADS43 and analyzed the expression of this gene in different tissues and shoot apical meristem periods.The results showed that the GhMADS43 gene was 696 bp in length and encoded 231 amino acids, which was similar to FUL in other plant species. According to specific expression analysis, GhMADS43 gene had higher expression in roots and apical buds, and the expression of the gene was higher in infinite fruit branches cotton CCRI50 than that of one fruit branches cotton Zaoling No.1.We also constructed bait vectorsp pGBKT7-GhMADS43 and transformed it into the yeast strain Y2HGold. The results showed that all the Y2HGold strains containing the recombinant plasmid grew well on the SD/-Trp media. It suggested that the recombinant bait proteins were no toxic to yeast. The transformed yeast strains developed blue plaques on the SD/-Trp/X-α-Gal media. And they could not grow on the SD/-Trp/-Ade/-His media with different concentration of 3-AT. The results indicated that the GhMADS43 gene had no self-activating activity. The constructed bait vector can be used for further screening of proteins interacting with GhMADS43 from yeast two-hybrid cDNA library in cotton. The study plays the foundation for analyzing of the gene function and regulatory pathways in shoot apical meristem and floral meristem development in the future.

Key words: Cotton, Meristem development, GhMADS43, Expression analysis, Self-activation

CLC Number:

Q78
S562.03

ZHANG Xiaohong, LI Xiaoqi, ZHANG Zheng, HU Genhai. Cloning and Yeast Self-activated Detection Analysis of GhMADS43 Gene in Cotton[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(6): 15-21. doi: 10.7668/hbnxb.20191274.

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