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Wiwit Budi Widyasari
Pusat Penelitian Perkebunan Gula Indonesia, Jl. Pahlawan 25 Pasuruan
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KONSTRUKSI VEKTOR BINER UNTUK EKSPRESI GEN dip22 (YANG DIISOLASI DARI TEBU VARIETAS M 442-51) PADA TANAMAN Wiwit Budi Widyasari; Sony Suhandono
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 13 No 1 (2007): December 2007
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/318

Abstract

Sugarcane is the principle plant for producing sugar in Indonesia. Water supply is one key element in the agronomy of sugarcane. Sugarcane is a high biomass crop which requires large amounts of water. Low yields of sugar observed in water stressed plants indicate that sugarcane is very sensititive to drought. A number of genes that respond to drought, salt, and cold stress at the trasnscriptional level have been reported. dip22 (drought inducible protein) protein isolated from drought resistance variety M 442-51 was predicted to be a protein regulator to water stress in sugarcane. Increasing of tolerance to water stress by over expression of dip22 genes in high yield sugarcane variety hopefully will maintain sugar production. The goal of this research was to construct a binary vector for dip22 gene expression in plant. dip22 gene from mutated PCR was cloned to pGEM®–T Easy and transformed to Escherichia coli strain DH5a. And then, these gene was isolated again from pGEM®–T Easy-dip22 (pGdip) plasmid using restriction enzymes NcoIand PmlI. pCAMBIA 1303 plasmid is an expression vector which has the constitutive promoter CaMV35S. Recombinant plasmid was transformed to Escherichia coli strain DH5a for plasmid propagation through DNA replication. Recombinant plasmid was isolated, and digested with NcoI and PmlI to examine the presence of dip22 gene in the pCAMBIA 1303 plasmid. The recombinant plasmid was transformed to A. tumefaciens strain LBA 4404. Plasmid isolated from A. tumefaciens was digested with Bst XI and Bst EII to examine the similarity between pCAMBIA 1303-dip22 (pCdip) from Escherichia coli and A. tumefaciens. The result by electrophoresis showed that both plasmids had the same size after digested. It was concluded that the transformed A. tumefaciens strain LBA 4404 bacteria has pCAMBIA 1303-dip22 (pCdip) plasmid indeed. Therefore, this construct of dip22 gene in binary vector can be used for improving drought tolerance in plant.
ISOLASI DAN ANALISIS GEN YANG RESPONSIF TERHADAP CEKAMAN KEKERINGAN PADA TEBU Wiwit Budi Widyasari; Bambang Sugiharto; Cahya Ismayadi; Kabul Wahjudi; Untung Murdiyatmo
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 9 No 2 (2004): June 2004
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/417

Abstract

This study was aimed to isolate and identify a gene that was responsive to drought stress in sugar cane variety M442-51, a variety that is widely used in pastures or dry land. Under drought stress, M442-51 showed an enhance expression of several proteins. One of the protein, namely dip22, (drought inducible protein, with a molecular weight of 22KDa) was isolated and analyzed. Amino acid sequence of this protein showed some homology with several stressed related proteins (OsAsr 1 from Oryza sativa leaves, ZM Bss1 of Zea mays leaves, LeAsr 1, LeAsr 2 and LeAsr 3 of Lycopersicum lycopersicon leaves). The 343 bp of dip22-cDNA fragment was obtained using RT PCR from mRNA of sugar cane leaves that has been exposed to drought stress. The results indicated that the gene dip22 plays an important role as a regulatory protein that are usually nested within cytoplasmic matrix or in the nucleus.
Co-Authors Bambang Sugiharto Cahya Ismayadi Kabul Wahjudi Sony Suhandono Untung Murdiyatmo