DNA isolation and identification of bioactive peptide sequences with anti- hypertensive activity in lemon grass (Cymbopogon citratus L.)

Date

4-2010

Degree

Bachelor of Science in Chemistry

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Mary Ann O. Torio

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Abstract

Search for bioactive peptides with antihypertensive activity in lemongrass was done using GenBank database. NADH dehydrogenase subunit F was found to contain peptide sequences LLP, FP, VF, LH, IYQY, IY, FY, VK, VW, LY and IR which are known to exhibit antihypertensive activity. Thus, primers specific for the DNA sequence containing these peptides were designed using VectorNTI and used for PCR amplification. Genomic DNA of lemongrass was extracted using the modified Cheung et al.- Datta et al.- Dellaporta et al. and the modified Doyle and Doyle extraction methods. Good quality genomic DNA was obtained using the modified Doyle and Doyle protocol, as evidenced by a single band in the agarose gel electropherogram. The isolated genomic DNA was used as a template for PCR. Results of PCR showed three major bands using the forward primer (5’-TCGCATATGGGCTTTTCC-3’) and the reverse primer (5’-CCTCTTCACGTATGGTTACCTG-3’). The bands correspond to products with 500 bp, 250 bp and 200 bp. However, bands corresponding to 500 bp was not purified and sent for DNA sequencing due to very low concentration. The other two bands, 250 bp (LG250) and 200 bp (LG200) sent for DNA sequencing revealed the presence of the ACE inhibitory peptides such as VW for LG200 and VW and IY for LG250 after translating the DNA sequence into peptides using Expasy Translate tool. Confirmation of the identity of the fragments amplified were done using UniProt BLAST search and BioEdit multiple alignment. Results from BLAST search showed that 247 of the 250 amino acid sequences for LG200 are NADH dehydrogenase subunit F of plants. However, no NADH dehydrogenase subunit F matched with the amino acid sequence of LG250.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2010 C4 R45

Document Type

Thesis

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