Analysis Of Expression And Function Of Low-temperature-induced Gene Wap25 Of Mulberry

There are different ecological types in germplasm resources of Mulberry, and every eco-types have different morphological characteristics. By using of mulberry trees in different eco-types and economic traits, it can not only guarantee the steady development of sericulture production, but also be applied to urban gardens and the ecological construction of sand-fixing. We chose germplasm which has resistance from mulberry resources, and cloned gene about cool, cold, drought, salt. We transfered low-temperature-induced protein from cold species into garden trees with weak cold hardiness to enhance frost resistance of trees in southern. It will provide wider value and more choice for garden plant and green project in cities. This paper research for low-temperature-induced protein gene of mulberry (Wap25) and gain some valuable parameters by using methods of bioinformatics. The results showed that Wap25 gene is one of the late embryonic-rich protein (LEA) genes. Compared with the other species, Wap25 gene is lower in homology. It consists of 226 amino acids, 25.3 kD molecular mass, 5.52 isoelectric point; N terminal 1-30 amino acid showed strong hydrophobicity, the following area showed high hydrophilicity with average value of -1.020. The 1-29 amino acid protein is typical eukaryotic proteins signal peptide sequence, it indecaded that WAP25 protein is secreted proteins.The target gene Wap25 was cloned into the prokaryotic expression vector pGEX-4T-2, and expressed in E. coli. successfully. And in vector pET-28a as the comparation in different IPTG concentrations, different time and different temperature conditions, the results showed there is no significant increase because of the low expression of target protein. Wap25 gene was cloned into the eukaryotic expression vector, then import the target gene to the onion epidermal cells by particle bombardment. Observated in the laser scanning confocal microscope , we found WAP25 protein located in the cytoplasm. Construction of RNA interference expression vector pRNAi-pA7-GFP-Wap25, we want to verify function of low-temperature-induced protein gene of Mulberry by gene gun technology.