Construction of yeast cDNA library and screening of TTM3 interaction protein in jujube fruit

Jujube fruit is rich in nutrition and contains a variety of bioactive substances such as cyclic adenosine monophosphate (cAMP), polysaccharide and vitamin C. The construction of a jujube fruit yeast cDNA library is an important premise to explore the molecular mechanism of jujube fruit development by yeast two-hybrid technology.
The jujube genome data show that the expression of TTM3 gene may be related to the efficient synthesis of cAMP in jujube fruit.
In this study, Gateway technology was used to construct the jujube fruit yeast cDNA library with pGADT7 as the carrier, using the white ripe and half-red fruits of ‘Dongzao’ as materials, and screening the proteins that interact with TTM3 through yeast two-hybridization.
The library quality test results showed that the library capacity was about 1.44×10⁷ CFU, the recombination rate was 96%, and the average length of the inserted cDNA fragments was greater than 1 kb.
The library was of high quality and met the requirements of yeast hybridization test.
The bait plasmid pGBKT7-PHYL1 was constructed, tested for its non-autoactivation activity, and transferred to Y2HGold yeast cells.
After yeast two-hybrid verification, 7 proteins that interact with TTM3 were screened out.
The construction of this library has important scientific significance for the study of jujube fruit development and the regulation mechanism of fruit nutrient components.

V International Jujube Symposium

Z.X. Zhao, Z.Q. Tian, Q. Niu, S.S. Cheng, Z.G. Liu, M.J. Liu, X. Zhao

jujube fruit, cDNA library, yeast two-hybrid, TTM3

https://doi.org/10.17660/ActaHortic.2022.1350.21