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- W2375846532 abstract "In order to elucidate the molecular regulation networks for soybean tolerance to abiotic stresses,a yeast two-hybrid cDNA library from soybean root tissue under high salt stress was constructed using SMART(Switching Mechanism at 5'End of the RNA Transcript) combined with DSN(Duplex-Specific Nulease) technique.The high-quality total RNA was isolated and subjected to reverse transcription using Oligo(dT)_(16),and the double-strand cDNAs were synthesized and further treated with DSN,then long fragment PCR amplification was performed using Advantage 2polymerase,electrophoresis results showed that the better homogeneous degree of PCR products ranged from 0.75-5.00 kb in length and the expression of GmActin significantly decreased by RT-PCR after homogenization.By using homologous recombination reaction,the dscDNAs were transformed into the Y187 yeast competent cell with the library plasmid pGADT7-Rec2 to construct the yeast two-hybrid cDNA library.Detection of the library quality demonstrated that it contained a total of 5.61×10~6 cfu independent yeast clones,the capacity of the library was up to 3.4×10~(10)cfu/mL,the recombinant rate was more than 90%,and the average size of inserts was approximately 1.0 kb in the cDNA library.These results demonstrated that the library meet the requirements for screening the interaction proteins and this will lay the foundation for insight into the soybean functional genomics" @default.
- W2375846532 created "2016-06-24" @default.
- W2375846532 creator A5042957435 @default.
- W2375846532 date "2013-01-01" @default.
- W2375846532 modified "2023-09-28" @default.
- W2375846532 title "Construction and analysis of a yeast two-hybrid cDNA library of soybean root tissue under high salt stress" @default.
- W2375846532 hasPublicationYear "2013" @default.
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