Mapping An Arabidopsis Plastid Transcriptional Active Chromosome Interaction Network

Pfalz separate35proteins of TACs(Transcriptional Active Chromosome) fromArabidopsis and Brassica juncea by mass-spectrography. Only some articles reportthe interaction of some proteins of TACs, the interaction of TACs complex have notbeen report. This article mainly study the interaction of TACs complex in Arabidopsisby yeast two-hybrid experiment. First, we determinate the candidate components ofTACs, the CDS of candidate components of TACs were cloned into the pGADT7vectorand pGBKT7vector.(The leader peptide of these genes are deleted.)Then, we mix thepGADT7plasmids which contain the CDS of candidate components of TACs as a pool.We will select positive clones for every bait gene, then amplified the insert ofplasmids cohabiting in each positive clone. Using the selection strategy, there are140interactions among30components. AtMurE (At1g63680), rpoB(Atcg00170),rpoC1(At00180), pTAC2(At1g74850), pTAC3(At3g04260) have no any proteininteraction with them. We find that Ptac6, Ptac12, Ptac14, FSD2, FSD3, FLN1, FLN2,TrXZ had more interaction proteins than other proteins. In addition, the T-DNAinsertion mutants of those genes exhibit albino or yellow cotyledon. At last, we drawa visualize picture of interaction of TACs complex. Here, we investigated the T-DNAknockout lines of ECB1which displayed albino phenotype on MS medium withoutsucrose, and decreased PEP-dependent plastid gene expression. The function ofECB1is unknown. Thus we construct the vector of yeast two-hybrid experiment ofECB1to bait the pool of TACs complex of Arabidopsis. We find that ECB1interact withpTAC9，TrXZ，pTAC18，FSD3，FLN1, this result set the basis for unraveling themolecular requirement for searching the function of ECB1.