Related Experiment Video
Updated: Jan 24, 2026

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
Published on: April 1, 2016
Combined Assembly and Targeted Integration of Multigene for Nitrogenase Biosynthetic Pathway in Saccharomyces
Xiaomeng Liu1, Minyang Wang1, Yi Song1
1State Key Laboratory for Agrobiotechnology, College of Biological Sciences and Key Laboratory of Soil Microbiology of Agriculture Ministry , China Agricultural University , Beijing , P. R. China.
Abstract:
Biological nitrogen fixation, a process unique to diazotrophic prokaryote, is catalyzed by the nitrogenase complex. There has been a long-standing interest in reconstituting a nitrogenase biosynthetic pathway in a eukaryotic host with the final aim of developing N2-fixing cereal crops. In this study, we report that a nitrogenase biosynthetic pathway (∼38 kb containing 15 genes) was assembled in two individual one-step methods via in vivo assembly and integrated at δ and HO sites in Saccharomyces cerevisiae chromosome. Of the 15 genes, 11 genes (nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA, nifV, groES, groEL) were from Paenibacillus polymyxa WLY78 and 4 genes (nifS, nifU, nifF, nifJ) were from Klebsiella oxytoca. The 15-gene nitrogenase biosynthetic pathway was correctly assembled and transcribed in the recombinant S. cerevisiae. The NifDK tetramer with an identical molecular weight as that of P. polymyxa was formed in yeast and the expressed NifH exhibited the activity of Fe protein. This study demonstrates that it will be possible to produce active nitrogenase in eukaryotic hosts.
Related Concept Videos
Amino Acid Biosynthetic Pathways
C4 Pathway and CAM
C4 Pathway
The C4 pathway is used by plants such as...
Protein Complex Assembly
Many viruses self-assemble into a fully functional unit using the infected host cell to...
Other Glycolytic Pathways
Respiration Pathways
Auditory Pathway
When viewed cross-sectionally, the cochlea reveals the scala vestibuli and scala tympani flanking...

