Homology search and purification of cel 1 homolog in tomato (Solanum lycopersicum L.)
Date
4-2014
Degree
Bachelor of Science in Agricultural Biotechnology
College
College of Agriculture and Food Science (CAFS)
Adviser/Committee Chair
Hayde F. Galvez
Abstract
CEL 1 is the used endonuclease for mutation cutting in TILLING (Targetting Induced Local Lesions IN the Genome). Homology search of CEL 1 was done in SOL Genomics Network (SGN) and National Center for Biotechnology Information (NCBI) to identify alternative Solanum sources for endonuclease. ENDO 1 gene of tomato has the highest homotology which is 74 % among all the Solanum species. Using SGN tools, it was found out that ENDO 1 is 2.182 kb gene containing eight introns and nine exons, and was classified under the endonuclease class. For molecular confirmation, primers were generated in Primer3.Amplification and specificity of the primers were checked using In silico PCR in SGN. Four primers were used for the amplification of ENDO 1 in five tomato samples, together with two rice samples. Results confirmed that ENDO 1 is present among five tomato samples. Also, CEL 1 homolog was confirmed to be present in rice. The purification of the expressed protein was done using the protocol for celery with some modifications. The purified ENDO 1 had the same mutation cutting specificity as compared to the stock CEL 1.
Language
English
Location
UPLB Main Library Special Collections Section (USCS)
Call Number
LG 993.5 2014 A127 A53
Recommended Citation
Alcasid, Rochelle Escobin, "Homology search and purification of cel 1 homolog in tomato (Solanum lycopersicum L.)" (2014). Undergraduate Theses. 2509.
https://www.ukdr.uplb.edu.ph/etd-undergrad/2509
Document Type
Thesis