Biotechnology Bulletin ›› 2017, Vol. 33 ›› Issue (12): 99-107.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0408

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Screening Interacted Factors of ZmSCL1 in Yeast Two-hybrid cDNA Libraries of Developing Maize Seeds

ZHAO Qian-qian, ZHOU Xiao-jin, CHEN Ru-mei   

  1. Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081
  • Received:2017-05-30 Online:2017-12-25 Published:2017-12-21

Abstract: ZmSCL1 is a member of GRAS gene family. Although highly expressed in maize seeds in the key period for the filling period of maize,especially in the endosperm,the molecular function of ZmSCL1 for maize seeds development remains unclear. Therefore,to screen interaction factors for ZmSCL1,yeast two-hybrid cDNA libraries of maize inbred line(B73)developing seeds(12 and 20 days after pollination)were constructed in yeast strain AH109 using homologous recombination-mediated SMART technology. The qualities of the cDNA libraries were evaluated as the followings:the titers were 1.10×105 CFU/mL,the capacity of cDNA library was 1.32×106,the recombinant rate was 81.90%,and the protein length encoded by the inserted cDNA ranged from 133 aa to 500 aa. Finally,109 clones were obtained for sequencing. GO enrichment analysis showed that,ZmSCL1 may be involved in the regulation of starch and nutrition accumulation.

Key words: GRAS gene family, SMART technology, cDNA library, ZmSCL1 gene