Isolation, amplification and characterization of DNA encoding hypocholesterolemic peptide from major storage protein of chickpea (Cicer arietinum L.) and peanut (Arachis hypogaea)

Date

5-2011

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Mark Rickard N. Angelia

Abstract

The presence of hypocholesterolemic peptide in the amino acid sequences of chickpea and peanut was determined by in silica analysis using the Basic Local Alignment Searching Tool (BLAST). Chickpea legumin was found to contain peptides EK, WK and IAVP while the peanut arachin showed the presence of the peptide IAVP. Primers which are specific to the hypocholesterolemic peptide of interest were designed using OligoCalc. Genomic DNA of chickpea and peanut were isolated using the method of Cheung et al., Datta et al. and Dellaporta et al. as modified by Angeles et al. (2005) and were used as template for polymerase chain reaction. Using the forward (5'. GGTATTCCCTGGTTGIGTTG-3') and the reverse printers (5'. GCTGCTGGTGICAATAAGAG-3'), a PCR product with 200 by was obtained for chickpea. For peanut, a PCR product with molecular weight corresponding to 250 by was amplified using the designed forward (5'-ACCAAAGCCAACAGCAACAG-3') and reverse primers (5'4TGCTTAGGCTGAGITTGCG-3'). The amplified products coded as Ch (chickpea) and PN (peanut) were sent to Macrogen for DNA sequencing analysis. Analysis of the sequence of Ch revealed the presence of the hypocholesterolemic peptide IAVP when translated to its amino acid counterpart using Expasy Translate tool. The sequence of PN, on the other hand, showed the presence of the peptides EK, VK and WK upon translation. These results confirmed that bioactive peptides exhibiting hypocholesterolemic activity are present in the protein sequences of chickpea and peanut.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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