Molecular cloning and characterization of small subunit ribulose-1, 5- bisposphate carboxylaseMolecular cloning and characterization of small subunit ribulose-1, 5- bisposphate carboxylase

Date

4-2012

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Lauena, Antonio C.

Abstract

A PCR-based approach of cloning the partial gene encoding the small subunit of Rubisco (rbcS) in malunggay (Moringa oleifera Lam.) using primers (RGI-RGIII) derived from conserved regions of the rbcS gene from several plant sources generated two amplicons, 850 and 500bp. The 500bp PCR fragment is the dominant amplicon and selected for cloning using the pGEM® T-Easy vector cloning system. After ligation, transformation, subsequent culture of transformed bacteria and plasmid miniprep, the recombinant plasmid was sent to an outside service facility for DNA sequencing. DNA sequence analysis of the 500bp PCR amplicon yielded at least two types of rbcS gene in malunggay, in terms of restriction sites, intron/exon size and BLAST search. There were subtle differences in the codon preference of malunggay as compared to other plant species. The two genes- rbcS1 and rbcS2 belong to the typeII rbcS based on the gene organization. The degenerate primers used for PCR-based gene cloning failed to generate polymorphic bands for DNA finger printing and diversity analyses of malunggay collected from different geographical locations in the Philippines. However, based on the DNA sequence data showing differences in the intron/exon size, new primers can be designed to produce polymorphic bands or amplicons. This is the first study to clone the partial gene of the malunggay rbcS gene.

Language

English

Location

UPLB Main Library Special Collections Section

Call Number

Thesis

Document Type

Thesis

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