CONTRUCTION AND TRANSIENT EXPRESSION OF DEHYDRIN PROMOTER FROM SUGARCANE (Saccharum officinarum L)

• Main Author: EKA PRASETYO (NIM : 10613049), AGUS
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/20817
• Institution: Institut Teknologi Bandung

Sugarcane plant is a plant that has a high level of tolerance to environmental stress. Dehydrin genes are the most widely regulated inducible stress genes in sugarcane plants compared to other inducible stress genes. The gene is controlled by promoter that has cis regularoty element in response to biotic and abiotic stress induction. This study aims to insert a dehydrin promoter from sugarcane into a pBI121 expression vector replacing the CaMV 35S promoter with Overlap Extention Polymerase Chain Reaction (OE-PCR) method and GUS assay in transient transformation. The OEPCE method requires a primary design that can amplify the overall pBI121 expression vector. The Dpn1 restriction enzyme was added to degrade the parental plasmid after the PCR Overlap process. The product was transformed to competent Escherechia coli DH5α. The promoter characterization was made transiently using Agrobacterium tumefaciens GV 3010 bacteria into tobacco tissue culture (Nicotiana tobacco Linn). Based on PCR confirmation, it has been shown that the plasmid pBI121-Pr1DHN1 has been successfully constructed into pBI121 expression vector and visualized with electrophoregram visualization on band of 2000 bp. The sequensing result was compared with the previous data show 99 % percen idendity with alignment tools BLASTn. The result of histochemical GUS staining assay showed that DHN11 promoters could regulate GUS gene expression in the stem. Esspcecially in xylem or floem. The low activity of the promoter dehydrin might the study did’nt include drought induction.