Cloning And Functional Characterization Of F128 And F103 Gene From Millet (Stetaria Italica)

Potato SB40J cDNA were used to screen the cDNA expression library of millet, and five clones were gained and sequenced. According to the results of Blast in GenBank+EMBL+DDBJ, three clones share high identity with genes that function is clearly. F128 and F103 share 56% amino acid sequence identity with Oryza saliva P0003D09.9 gene, 34% with Arabidopsis thaliana At2g42190 gene separately. We choose F128 and F103 for further research.F128 gene is composed of 581 nt. It was deduced that it contains an open reading frame coding for a ll,691Da polypeptide of 109 amino acid residues. The results of analysis by Biology Software were that in the structure of the deduced protein the hydrophobic domain is dominant, and there are several phospho-sites. It seems to have a cleavable N-term signal seqence,and it may be a vacuolar protein.Genomic Southern blot analysis indicated that F128 mRNA is encoded by a single gene in millet. Northern blot analysis showed that F128 mRNA accumulates in developing seeds. Genomic Southern DNA analysis of F128 homologous gene in some plants showed that F128 has homologous gene in maize.Plant expression vector pBO128S were constructed and introduced into tobacco in order to study the function ofF128. The phenorype difference of transformed tobacco from control is that callus develops three to four days later, but the shoot is close to control.F103 gene is composed of 819 nt. It was deduced that it contains an open reading frame coding for a 14,113 Da polypeptide of 130 amino acid residues. The results of analysis by Biology Software were that in the structure of the deduced protein the hydrophibic domain is dominant, and there are several phospho-sites. It may be a nuclear targeting protein.Genomic Southern blot analysis indicated that F103 mRNA is encoded by a single gene in millet. Northern blot analysis showed that F103 mRNA accumulates in stem, inflorescence, developing seeds, but not in leaves. Genomic Southern DNA analysis of F103 homologous gene in some plants showed that F103 has homologous gene in maize.Plant expression vector pBO103S were constructed and introduced into tobacco in order to study the function of F103. The phenorype difference of transformed tobacco from control is that callus develops one week later, but the shoot is close to control.