Cloning And Expression Analysis Of ZFP-1 Gene Associated With The Flower Tube Cracking From Lonicera Macranthoides

In this paper, based on the sequencing of transcription, the full length cDNA of ZFP-1 with the structure of the zinc finger from Lonicera macranthoides flower tube were cloned with RT-PCR and RACE, and the bioinformatics analysis, fluorescence quantitative expression for ZFP-1 gene were done. By the gene expression patterns in the different periods of floral organ development, suggesting that this gene plays a role in the development of floral organs, set up a foundation for the molecular regulation mechanism of flower tube splitting, and provide a theoretical basis for molecular breeding. The results were as follows:(1) The full length cDNA of ZFP-1gene was synthesized using RT-PCR and RACE. The gene was 1647bp in length with an open reading frame (ORF) encoding 371 amino acids, a 5’un-translated region of 326bp and a 3’flanking sequence of 205bp. The sequences were submitted to GenBank (Accession number KRO13240). Compared with ZFP gene of others plants by Blast software, ZFP-1 gene respectively had 77%,77%,76%,76% and 75%identity with that of Prunus mume, Prunus persica, Theobroma cacao, Malus and Citrus.(2) ZFP-1 gene of Lonicera macranthoides belong to the zinc finger protein gene families, a BTB/PZO domain at the N end and a zf-TAZ domain at the C end. Results of BLAST and amino acid sequence alignment showed 75% amino acid sequence homology with Theobroma cacao,74% with Vitis vinifera.(3)The molecular weight of ZFP-1 was determined as 4.28kD with 46.1% non-polar amino acid,24.5% uncharged polar amino acid,18.1% basic amino acid, 11.3% acidic amino acid. The isoelectric point (p1) of the gene was found at 9.18. The number of polar amino acid is slightly higher than the number of non-polar amino acid. It can be speculated that the ZFP-1 protein is a hydrophilic protein.(4)The main secondary structure of the ZFP-1 protein is curl and spiral structure. The existence of a trans-membrane structure near N end, is located in the 104～122, the length of 19 amino acids. There is also a signal peptide, amino acids located at 1-70.(5)The qRT-PCR analysis showed enhanced transcript number of ZFP-1 gene with increasing the stage of flower bud development. The expression of the ZFP-1 gene in’putong’is very low, almost no expression, but highly expression in the ’jincuilei’.