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
Recommended Citation
Mercado, Mark Julius B., "Screening and characterization of filtered genomic DNA library from sugarcane (Saccharum officinarun L.) var. VMC 86-550 using methylation-sensitive restriction enzyme" (2016). Undergraduate Theses. 4889.
https://www.ukdr.uplb.edu.ph/etd-undergrad/4889
Document Type
Thesis
Notes
Major in Crop Biotechnology