Identification And Prokaryotic Expression Of A Gene Encoding C3HC4-type Ring Zinc Finger Protein From Dongxiang Wild Rice

Dongxiang wild rice is the unique common wild rice found in the most northern ofthe world that can survive overwintering so far. It contains a wealth of superiorgermplasm resources, such as cold-, disease-, and insect-resistance and so on. Soidentification and utilization of genes of high quality traits from Dongxiang wild rice isof great significance. In this work, a gene RZF with an ORF of705bp encodingC3HC4-type RING zinc finger protein was screened and identified from cDNA library ofthe Dongxiang Wild rice. Bioinformatic analysis of the protein showed that this RINGzinc finger protein is mainly composed of polar amino acids exhibiting99%homologywith that of common rice. Structural analysis manifested that this protein contained atransmembrane region and a ring zinc finger domain which putatively providing forprotein-protein interactions. By analyzing the upstream promoter sequence of the gene, avariety of cis-elements that can be bound by proteins such as ABRELATERD1,CBFHV，MYB1AT were found, thus the gene of C3HC4-type RING zinc finger proteinwas predicted involved in the signal pathway induced by ABA and dehydration.Besides，we construct the recombinant vector pET28-RZF, then express induced bythe IPTP. SDS-PAGE electrophoresis detection shows that the expressed protein has aapproximate molecular weight of30.0kD consistent with the expected size. Thepurification of protein can obtain good results by using Ni-NTA Superflow column. Inaddition, this paper utilizate the relationship between gene and cold stress, the expressionin different mature tissues, as well as the structure and function of this protein to predict.This work has laid a good foundation for the function identification of C3HC4type ringzinc finger protein in wild rice.