Construction and Transient Analysis of Metuf C Promoter from Cassava (Manihot esculenta Crantz.) Var. Adira in Tobacco Plant (Nicotiana tabacum L.)

• Main Author: RIZKIA (NIM : 10613016), ANNISA
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/21149
• Institution: Institut Teknologi Bandung

Metuf C promoter is a promoter which regulates expression of Metuf C gene. Expression of this gene generates EF-Tu (Elongation Factor Thermo unstable) protein. EF-Tu plays an important role as translation elongation factor also has chaperone activity. EF-Tu increase under heat-induced condition. One of promoter which belongs to Metuf gene family called Metuf C has HSE (Heat Stress Element). This element supports plant tolerance to heat stress. The study aims to construct Metuf C promoter and analyze it with transient method in tobacco plants. Metuf C promoter construct has been made using Overlap Extension PCR Cloning (OEPC) technique. Mega primer was generated by combining insert and pBI121 overlap sequence to replace CaMV35S promoter. OEPC product was transformed to Escherichia coli DH5α vector. PCR confirmation showed that band which has 1268 bp size appear on the sample. Sequencing analysis with pairwise sequence alignment generate 99% score percent identity with Metuf C promoter. Construct was analyze in Agrobacterium tumefaciens strain AGL1 with agroinfiltration method. According to histochemical GUS Assay, promoter Metuf C element regulates expression of GUS gene in tobacco root, stem, and leaf. So, this study suggest Metuf C is a functional promoter that can be a candidate to regulate expression of gene in crop plants.