The Superman Gene Cloning And Preliminary Analysis In Asparagus Officinalis

Plant SUPERMAN zinc finger protein belongs to the C2H2type and has a single zinc finger motif. Ithas a N-terminal consensus sequence of QALGGH and a C-terminal consensus sequence ofL(I)DLXLR(K)L, and it mainly exerts its function during the development of plants' flower, lateral root,seed and embryo. Recent study showed that the SUPERMAN gene may participate in the regulation of sexdetermination in dioecious plants. In order to research the SUPERMAN genes are involved in the sexdetermination and the differentiation process of Asparagus officinalis, using dioecious plant Asparagusofficinalis as subjects and according to the reported consensus sequence of SUPERMAN zinc finger proteingene, we designed primers and cloned a SUPERMAN zinc finger protein gene named AoSUP and madebrief analysis on its gene sequence, protein structure, differentiated expression in certain vegetative tissueof male and female plants. The results were as follows.1, The AoSUP full length cDNA sequence was cloned with the rapid amplification of cDNA ends(RACE). The sequence analysis showed that AoSUP has a1052bp full length sequence, which include a409bp conserved sequence and a786bp open reading frame (ORF). This ORF encoding a261amino acidspeptide (the5'-UTR is63bp and the3'-UTR is213bp). Amino-acid sequence alignments between AoSUPand11other typical plant SUPERMAN zinc finger gene, and the NJ evolutionary tree showed that AoSUPhas a15%-49%similarity with them. And of the11SUPERMAN single zinc finger proteins, the KUN geneof Arabidopsis is the most similar, accounting for49%; its amino acid sequence contain a core zinc fingerlike domain QALGGHM*R*R, and a C-terminal L/IDLELRLG sequence.2, The AoSUP amino acid sequence was analysed with the software including ProtParam, ProtScale,SOPMA and NetPhos2.0Serve, the results showed that AoSUP molecular weight is28361.0, pI is10.16,and has more hydrophilic than hydrophobic amino acids, thus it is a hydrophilic protein. The secondarystructure of AoSUP contains mainly α-helix, β-sheet and β-turn, accounting for27.20%,12.64and5.36respectively. AoSUP contains17phosphorylation sites in total, including11Ser sites,3Thr sites and3Tyrsites.3, The expression pattern analysis of the AoSUP gene in male and female plants was usedRT-PCR.The results showed that AoSUP is both expressed in male and female Asparagus officinalis's root, stem, leaf and flower, with lowest expression found in root, the stem and leaf and flower of the expressionis identical. No significant difference of AoSUP expression was observed between male and female plants. Sowe assumed that AoSUP may not be taken part sex determination and differentiation process in Asparagusofficinalis's.