Extraction, detection,and characterization of free nucleic acid-polysaccharide (FNA-PS) complex from the water of macapuno coconut (Cocos nucifera L.).

Date

4-2006

Degree

Bachelor of Science in Biology

Major Course

Major in Cell Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Marni E. Cueno

Abstract

To determine the presence of free nucleic acid (FNA) in the water of makapuno coconut (Cocos nucifera L.), the extraction protocol for FNA in makapuno water was optimized using a volume of 10 mL of makapuno water and three salts: sodium acetate, potassium acetate and ammonium acetate. Detection of FNA using agarose gel electrophoresis was not possible in all set-ups. However, detection of FNA using colorimetric test was successful in all three set-ups, suggesting FNA exists and forms a complex with high molecular weight molecules. To prove this association, a modified Michaels et al. (1994) protocol was employed. The extracted polysaccharides (PS) were positive in the colorimetric test confirming association of PS with FNA. Sodium acetate gave the highest concentration of nucleic acid with 1 ng/uL, proving to be the most efficient among the three salts. The pattern of FNA-PS association was determined thru Proteinase K digestion. A considerable high amount of FNA detected in the isolated polysaccharide after protein digestion confirms the presence of FNA-PS complex. Characterization of FNA-PS complex was done thru DNase I digestion and RNase A digestion. Results showed a higher concentration of RNA (5 ng/ uL) compared to DNA (0.5 ng/uL) in makapuno water. These findings would suggest the probable role of FNA-PS (RNA-PS) complex in the regulation of enzyme activity. Defining the structural association of FNA-PS complex would broaden the knowledge of the mechanism of interaction of the two. Furthermore, the results would set foundation for future molecular biology researches with makapuno water.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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