Screening and characterization of filtered genomic DNA library from sugarcane (Saccharum officinarun L.) var. VMC 86-550 using methylation-sensitive restriction enzyme

Date

6-2016

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Antonio C. Laurena

Restrictions

Restricted: Not available to the general public. Access is available only after consultation with author/thesis adviser and only to those bound by the confidentiality agreement.

Abstract

The use of methylation-sensitive restriction enzyme, PstI, in cleaving functional genomic DNA was successful. Moreover, the sequences of the 30 cloned DNA fragments were unique and indicating zero redundancy. Among the 30 sequences analyzed, (37.5%) were biosynthesis-related genes of various functions, (9.4%) were clones from Saccharum hybrids, (46.9%) were uncharacterized proteins and (6.2%) were clones with no hits. Clones that were aligned to the Saccharum cultivars have the highest similarity with 100%. One clone, PstI-4, was aligned to a methionyl-tRNA synthethase that is essential for protein biosynthesis. The clone PstI-6 was aligned to a jacalin-like plant lectin, a protein that binds to sugar. Another clone, PstI-10, was aligned to a cinnamic acid 4-hydroxylase that is a major part of lignin biosynthesis. Clone PstI-7 aligned to a permease that has an effect on sugar production. Moreover, two novel sequences PstI-14 and PstI-34 have been cloned for from the methylation filtered genome of sugarcane variety VMC 86-550. A tetranucleotide repeat was also detected in the nucleotide sequence clone PstI-7 and SSR primers were designed to amplify the microsatellite region for testing. Results of the study have demonstrated the utility of PstI in the construction of methylation-filltered genomic DNA library for gene and DNA marker discovery.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2016 A127 /M47

Notes

Major in Crop Biotechnology

Document Type

Thesis

This document is currently not available here.

Share

COinS