Purification and Gene Expression Profiling of Lipase from Oryza sativa

Poornima, Dubey (2010) Purification and Gene Expression Profiling of Lipase from Oryza sativa. [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type:	Student Project Report
Additional Information:	The partial purification of rice bran lipase was done from the rice flour sample The specific activity of crude sample was found to be 032U/mg. After gel filtration chromatography the specific activity was increased to 0.1299U/mg and increase in total purification of lipase was found to be 4.0312 folds. The Native PAGE gel was run and after activity staining activity bands were observed. Temporal expression profiling of rice lipase was done by germinating rice seeds for different time intervals. The activity of lipase protein expressed at different stages of germination was checked and it was found to be maximum between 4th to 7th days of germination. The germinated samples of different stages were taken and RNA was isolated. The total RNA was isolated by Trizol method and Spectrophotometric reading shows ratio of A260/A280 1.92. Formaldehyde gel was run and bands were observed from samples of different time intervals. cDNA of isolated RNA was prepared by RT PCR. The presence of amplified cDNA was checked by using actin specific primers and actin’s cDNA was act as positive control because actin is a housekeeping gene and amplicons were observed for actin’s cDNA. The genomic DNA was isolated from rice flour (IR 64 variety) and the 143bp RBL primers were used for amplification of lipase DNA fragments of 143bp.and gel was run and band was observed. Further attempts are in progress in this laboratory.
Uncontrolled Keywords:	rice bran lipase; rice flour; Temporal expression profiling
Subjects:	600 Technology > 08 Food technology > 21 Cereals > 01 Rice 500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 16 Enzyme Chemistry
Divisions:	Protein Chemistry and Technology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	21 Jul 2010 06:00
Last Modified:	28 Dec 2011 10:16
URI:	http://ir.cftri.res.in/id/eprint/9586

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