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

Document Type

Thesis

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